82,136 results on '"Nuclear Proteins"'
Search Results
2. 3-Nitropropionic Acid Enhances Ferroptotic Cell Death via NOX2-Mediated ROS Generation in STHdhQ111 Striatal Cells Carrying Mutant Huntingtin
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Mice ,Huntington Disease ,Caspases ,Animals ,Nuclear Proteins ,Neurodegenerative Diseases ,Mice, Transgenic ,Nerve Tissue Proteins ,Reactive Oxygen Species - Published
- 2023
3. Recurrent KAT6B/A::KANSL1 Fusions Characterize a Potentially Aggressive Uterine Sarcoma Morphologically Overlapping With Low-grade Endometrial Stromal Sarcoma
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Abbas Agaimy, Blaise A. Clarke, David L. Kolin, Cheng-Han Lee, Jen-Chieh Lee, W. Glenn McCluggage, Patrik Pöschke, Robert Stoehr, David Swanson, Gulisa Turashvili, Matthias W. Beckmann, Arndt Hartmann, Cristina R. Antonescu, and Brendan C. Dickson
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Adult ,Aged, 80 and over ,Leiomyoma ,Sarcoma, Endometrial Stromal ,Nuclear Proteins ,Soft Tissue Neoplasms ,Middle Aged ,Pathology and Forensic Medicine ,Endometrial Neoplasms ,Endometrial Stromal Tumors ,Uterine Neoplasms ,Biomarkers, Tumor ,Humans ,Surgery ,Female ,Neprilysin ,Anatomy ,Aged ,Histone Acetyltransferases - Abstract
With the widespread application of next-generation sequencing, the genetic landscape of uterine mesenchymal neoplasms has been evolving rapidly to include several recently identified fusion genes. Although chromosomal rearrangements involving the 10q22 and 17q21.31 loci have been reported in occasional uterine leiomyomas decades ago, the corresponding KAT6B::KANSL1 fusion has been only recently identified in 2 uterine tumors diagnosed as leiomyoma and leiomyosarcoma. We herein describe 13 uterine stromal neoplasms carrying a KAT6B::KANSL1 (n=11) and KAT6A::KANSL1 (n=2) fusion. Patient ages ranged from 33 to 81 years (median, 49 y). Tumor size was 2.6 to 23.5 cm (median, 8.2 cm). Nine tumors were myometrium-centered, and 3 had an intracavitary component. Original diagnoses were mostly low-grade endometrial stromal sarcoma (LG-ESS; 10 cases) with atypical features (limited CD10 expression, sex cord-like features, pericytic vasculature, and frequent myxoid changes). Treatment was hysterectomy±bilateral salpingo-oophorectomy (10), myomectomy (1), and curettage (2). Five patients were disease-free at 6 to 34 months, 3 (27%) died of disease at 2 to 47 months, and 3 were alive with disease at 2, 17, and 17 years. Histologically, most tumors showed variable overlap with LG-ESS, but they were generally well-circumscribed lacking the extensive permeative and angioinvasive growth typical of LG-ESS. They were composed of monotonous medium-sized oval and spindle cells arranged into diffuse sheets with prominent spiral-type arterioles and frequent pericytoma-like vascular pattern. Variable myxoid stromal changes were frequent. Mitotic activity ranged from 1 to20 in 10 HPFs. Immunohistochemistry showed variable expression of CD10 (12/13), estrogen receptor (8/11), progesterone receptor (8/11), smooth muscle actin (9/11), desmin (4/12), h-caldesmon (2/10), calretinin (3/8), inhibin (1/7), WT1 (4/7), cyclin D1 (5/11; diffuse in only 1 case), and pankeratin (5/10). This series characterizes a KAT6B/A::KANSL1 fusion-positive uterine stromal neoplasm within the morphologic spectrum of LG-ESS but with atypical features. The relationship of these neoplasms to genuine LG-ESS remains unclear. This molecular subtype of uterine endometrial stromal sarcoma has the potential for an unfavorable clinical course despite the absence of widely invasive growth; nevertheless, analysis of more cases is necessary to delineate the phenotypic spectrum and biological potential of this tumor.
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- 2023
4. Targeted degradation via direct 26S proteasome recruitment
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Charlene Bashore, Sumit Prakash, Matthew C. Johnson, Ryan J. Conrad, Ivy A. Kekessie, Suzie J. Scales, Noriko Ishisoko, Tracy Kleinheinz, Peter S. Liu, Nataliya Popovych, Aaron T. Wecksler, Lijuan Zhou, Christine Tam, Inna Zilberleyb, Rajini Srinivasan, Robert A. Blake, Aimin Song, Steven T. Staben, Yingnan Zhang, David Arnott, Wayne J. Fairbrother, Scott A. Foster, Ingrid E. Wertz, Claudio Ciferri, and Erin C. Dueber
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Ligases ,Proteasome Endopeptidase Complex ,Ubiquitin-Protein Ligases ,Cryoelectron Microscopy ,Proteolysis ,Nuclear Proteins ,Cell Biology ,Molecular Biology ,Transcription Factors - Abstract
Engineered destruction of target proteins by recruitment to the cell’s degradation machinery has emerged as a promising strategy in drug discovery. The majority of molecules that facilitate targeted degradation do so via a select number of ubiquitin ligases, restricting this therapeutic approach to tissue types that express the requisite ligase. Here, we describe a new strategy of targeted protein degradation through direct substrate recruitment to the 26S proteasome. The proteolytic complex is essential and abundantly expressed in all cells; however, proteasomal ligands remain scarce. We identify potent peptidic macrocycles that bind directly to the 26S proteasome subunit PSMD2, with a 2.5-Å-resolution cryo-electron microscopy complex structure revealing a binding site near the 26S pore. Conjugation of this macrocycle to a potent BRD4 ligand enabled generation of chimeric molecules that effectively degrade BRD4 in cells, thus demonstrating that degradation via direct proteasomal recruitment is a viable strategy for targeted protein degradation.
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- 2022
5. Loss of KDM5B ameliorates pathological cardiac fibrosis and dysfunction by epigenetically enhancing ATF3 expression
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Bo Wang, Yong Tan, Yunkai Zhang, Sheng Zhang, Xuewen Duan, Yuyu Jiang, Tong Li, Qingqing Zhou, Xingguang Liu, and Zhenzhen Zhan
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Heart Failure ,Jumonji Domain-Containing Histone Demethylases ,Activating Transcription Factor 3 ,Ventricular Remodeling ,Myocardium ,Clinical Biochemistry ,Nuclear Proteins ,Fibrosis ,Biochemistry ,Histones ,Repressor Proteins ,Humans ,Molecular Medicine ,Cyclic AMP Response Element-Binding Protein ,Molecular Biology - Abstract
Excessive cardiac fibrosis is central to adverse cardiac remodeling and dysfunction leading to heart failure in many cardiac diseases. Histone methylation plays a crucial role in various pathophysiological events. However, the role of histone methylation modification enzymes in pathological cardiac fibrosis needs to be fully elucidated. Here, we identified lysine demethylase 5B (KDM5B), a histone H3K4me2/me3 demethylase, as a key epigenetic mediator of pathological cardiac fibrosis. KDM5B expression was upregulated in cardiac fibroblasts and myocardial tissues in response to pathological stress. KDM5B deficiency markedly ameliorated cardiac fibrosis, improved cardiac function, and prevented adverse cardiac remodeling following myocardial infarction (MI) or pressure overload. KDM5B knockout or inhibitor treatment constrained the transition of cardiac fibroblasts to profibrogenic myofibroblasts and suppressed fibrotic responses. KDM5B deficiency also facilitated the transformation of cardiac fibroblasts to endothelial-like cells and promoted angiogenesis in response to myocardial injury. Mechanistically, KDM5B bound to the promoter of activating transcription factor 3 (Atf3), an antifibrotic regulator of cardiac fibrosis, and inhibited ATF3 expression by demethylating the activated H3K4me2/3 modification, leading to the enhanced activation of TGF-β signaling and excessive expression of profibrotic genes. Our study indicates that KDM5B drives pathological cardiac fibrosis and represents a candidate target for intervention in cardiac dysfunction and heart failure.
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- 2022
6. RanBP1 plays an essential role in directed migration of neural crest cells during development
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Elias H. Barriga, Delan N. Alasaadi, Chiara Mencarelli, Roberto Mayor, and Franck Pichaud
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Pregnancy ,Neural Crest ,Cell Movement ,Chemotaxis ,Humans ,Nuclear Proteins ,Female ,Cell Biology ,Molecular Biology ,Developmental Biology - Abstract
Collective cell migration is essential for embryonic development, tissue regeneration and repair, and has been implicated in pathological conditions such as cancer metastasis. It is, in part, directed by external cues that promote front-to-rear polarity in individual cells. However, our understanding of the pathways that underpin the directional movement of cells in response to external cues remains incomplete. To examine this issue we made use of neural crest cells (NC), which migrate as a collective during development to generate vital structures including bones and cartilage. Using a candidate approach, we found an essential role for Ran-binding protein 1 (RanBP1), a key effector of the nucleocytoplasmic transport pathway, in enabling directed migration of these cells. Our results indicate that RanBP1 is required for establishing front-to-rear polarity, so that NCs are able to chemotax. Moreover, our work suggests that RanBP1 function in chemotaxis involves the polarity kinase LKB1/PAR4. We envisage that regulated nuclear export of LKB1 through Ran/RanBP1 is a key regulatory step required for establishing front-to-rear polarity and thus chemotaxis, during NC collective migration.
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- 2022
7. Removal of nonspecific binding proteins is required in co-immunoprecipitation with nuclear proteins
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Xi Zeng, Wei-Hong Zeng, Jun Zhou, Xiao-Min Liu, Guo Huang, Hong Zhu, Shuai Xiao, Ying Zeng, and Deliang Cao
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Nuclear Proteins ,Immunoprecipitation ,Tumor Suppressor Protein p53 ,Carrier Proteins ,Actins ,General Biochemistry, Genetics and Molecular Biology ,Protein Binding ,Biotechnology - Abstract
Whether protein samples should be pretreated to remove nonspecific binding proteins in co-immunoprecipitation (CO-IP) is controversial. In this work, nonspecific binding of proteins to agarose beads was found to be greater than that to magnetic beads. The nonspecific binding was increased with the decrease of ion concentrations but reduced by Nonidet P40. Western blot indicated that p65 and β-actin were present as nonspecifically bound protein to the beads. p53 and β-actin were present in the CO-IP precipitates of nuclear proteins but pretreatment cleared the nonspecifically pulled down p53 and β-actin. These data suggest that magnetic beads are better for CO-IP, but preclearing is necessary to minimize false positive regardless of which bead is used, particularly for nuclear proteins.
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- 2022
8. Circ_0091579 exerts an oncogenic role in hepatocellular carcinoma via mediating miR-136-5p/TRIM27
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Yantao Mao, Zhigang Ding, Maozhu Jiang, Bo Yuan, Yao Zhang, and Xiaobin Zhang
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Carcinoma, Hepatocellular ,Liver Neoplasms ,Nuclear Proteins ,Apoptosis ,General Medicine ,Gene Expression Regulation, Neoplastic ,DNA-Binding Proteins ,Disease Models, Animal ,MicroRNAs ,Cell Line, Tumor ,Humans ,Animals ,Transcription Factors ,Cell Proliferation - Abstract
Circular RNAs (circRNAs) act as crucial regulators in tumorigenesis. In this study, the working mechanism of circ_0091579 in hepatocellular carcinoma (HCC) progression was investigated.The expression of RNA and protein was measured via RT-qPCR and Western blot assay. Cell proliferation ability was analyzed via CCK8, EdU and colony formation assays. Cell migration and invasion abilities were detected via transwell assays. Flow cytometry was applied to assess cell cycle and apoptosis. The target relation between miR-136-5p and circ_0091579 or tripartite motif containing 27 (TRIM27) was certified using dual-luciferase reporter assay. Xenograft tumor model was utilized to assess the role of circ_0091579 in tumor growth in vivo. The protein level of Ki67 in tumor tissues was analyzed by immunohistochemistry (IHC) assay.Circ_0091579 expression was elevated in HCC tissues and cell lines. HCC patients with high circ_0091579 expression displayed low survival rate. Circ_0091579 knockdown suppressed the proliferation, migration, invasion, cell cycle progression and epithelial-mesenchymal transition (EMT) and induced apoptosis of HCC cells. Circ_0091579 acted as a molecular sponge for miR-136-5p, and circ_0091579 silencing-mediated effects were largely overturned by the knockdown of miR-136-5p in HCC cells. MiR-136-5p interacted with the 3' untranslated region (3'UTR) of TRIM27, and TRIM27 overexpression largely counteracted miR-136-5p overexpression-induced influences in HCC cells. Circ_0091579 sponged miR-136-5p to up-regulate TRIM27 expression in HCC cells. Circ_0091579 silencing suppressed xenograft tumor growth in vivo.Circ_0091579 exhibited an oncogenic role to enhance the malignant potential of HCC cells through mediating miR-136-5p/TRIM27 axis in vitro and in vivo.
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- 2022
9. NOLC1 knockdown suppresses prostate cancer progressions by reducing AKT phosphorylation and β-catenin accumulation
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Wansoo Kim, Dong-Yeop Yeo, Seong-Kyoon Choi, Hee-Yeon Kim, Seoung-Woo Lee, Janbolat Ashim, Jee Eun Han, Wookyung Yu, Hyohoon Jeong, Jin-Kyu Park, and Song Park
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Male ,Prostate ,Biophysics ,Nuclear Proteins ,Androgen Antagonists ,Cell Biology ,Phosphoproteins ,Biochemistry ,Prostatic Neoplasms, Castration-Resistant ,Receptors, Androgen ,Cell Line, Tumor ,Humans ,Phosphorylation ,Proto-Oncogene Proteins c-akt ,Molecular Biology ,beta Catenin - Abstract
Although several studies have focused on cancer diagnosis and therapy, prostate cancer (PC) remains an intractable disease. Androgen deprivation therapy (ADT), which is used to treat early stage PC can lead to the development of castration-resistant prostate cancer (CRPC), which is highly associated with androgen receptor (AR) mutations. Nucleolar and coiled-body phosphoprotein 1 (NOLC1) is a chaperone that shuttles between the nucleus and the cytoplasm. Studies suggest that NOLC1 regulates PC progression; however, the underlying mechanisms remain unclear. Herein, we showed that NOLC1 knockdown suppresses PC cell proliferation by altering the signaling pathways and the expression of various proteins involved in DNA replication, amino acid metabolism, and RNA processing. Mechanistically, NOLC1 knockdown suppressed cell cycle progression by inhibiting AKT phosphorylation and β-catenin accumulation. Finally, we showed that NOLC1 expression is higher in human PC than in human hyperplastic prostate tissues. Altogether, we demonstrated that NOLC1 knockdown suppresses the progression of both AR-positive and AR-negative PC cells by inducing changes in the expression of several genes leading to cell cycle arrest. Thus, NOLC1 might be a novel and promising therapeutic target for PC.
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- 2022
10. MTR4 adaptor PICT1 functions in two distinct steps during pre-rRNA processing
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Sotaro Miyao, Kanako Saito, Renta Oshima, Kohichi Kawahara, and Masami Nagahama
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Saccharomyces cerevisiae Proteins ,Exosome Multienzyme Ribonuclease Complex ,Tumor Suppressor Proteins ,Oligonucleotides ,Biophysics ,Nuclear Proteins ,Saccharomyces cerevisiae ,Cell Biology ,Biochemistry ,RNA, Ribosomal, 5.8S ,RNA Precursors ,Humans ,RNA Processing, Post-Transcriptional ,RNA, Small Interfering ,Molecular Biology ,RNA Helicases - Abstract
Ribosome biogenesis proceeds with the successive cleavage and trimming of the large 47S rRNA precursor, where the RNA exosome plays major roles in concert with the Ski2-like RNA helicase, MTR4. The recent finding of a consensus amino acid sequence, the arch-interacting motif (AIM), for binding to the arch domain in MTR4 suggests that recruitment of the RNA processing machinery to the maturing pre-rRNA at appropriate places and timings is mediated by several adaptor proteins possessing the AIM sequence. In yeast Saccharomyces cerevisiae, Nop53 plays such a role in the maturation of the 3'-end of 5.8S rRNA. Here, we investigated the functions of PICT1 (also known as GLTSCR2 or NOP53), a mammalian ortholog of Nop53, during ribosome biogenesis in human cells. PICT1 interacted with MTR4 and exosome in an AIM-dependent manner. Overexpression of PICT1 mutants defecting AIM sequence and siRNA-mediated depletion of PICT1 showed that PICT1 is involved in two distinct pre-rRNA processing steps during the generation of 60S ribosomes; first step is the early cleavage of 32S intermediate RNA, while the second step is the late maturation of 12S precursor into 5.8S rRNA. The recruitment of MTR4 and RNA exosome via the AIM sequence was required only during the late processing step. Although, the depletion of MTR4 and PICT1 induced stabilization of the tumor suppressor p53 protein in cancer cell lines, the depletion of the exosome catalytic subunits, RRP6 and DIS3, did not exert such an effect. These results suggest that recruitment of the RNA processing machinery to the 3'-end of pre-5.8S rRNA may be involved in the induction of the nucleolar stress response, but the pre-rRNA processing capabilities themselves were not involved in this process.
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- 2022
11. Primary cutaneous SMARCA4-deficient undifferentiated malignant neoplasm: first two cases with clinicopathologic and molecular comparison to eight visceral counterparts
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Eleanor Russell-Goldman, Laura MacConaill, and John Hanna
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Neuroblastoma ,Biomarkers, Tumor ,Carcinoma, Squamous Cell ,DNA Helicases ,Humans ,Nuclear Proteins ,Sarcoma ,Prognosis ,Immunohistochemistry ,Transcription Factors ,Pathology and Forensic Medicine - Abstract
SMARCA4-deficient undifferentiated malignant neoplasms (SD-UMN) comprise a group of aggressive tumors with epithelioid morphology that are characterized by loss of function of SMARCA4, a component of the SWI/SNF chromatin remodeling complex. SD-UMN was first recognized in the thoracic cavity but is now appreciated to occur at multiple anatomic sites. A notable exception has been skin. Here we report the first two cases of primary cutaneous SD-UMN and compare their features to a cohort of eight visceral cases arising in lung, gastrointestinal tract, and gallbladder. Evidence for a bona fide cutaneous origin included extensive clinical, radiologic, and serologic analyses that failed to identify a metastatic source as well as the molecular identification of a UV-associated mutational pattern. The cutaneous cases showed strikingly similar morphologic, immunohistochemical, and molecular features to the visceral cases, strongly suggesting that they belong to this family of tumors. In addition to biallelic inactivation of SMARCA4, both cutaneous tumors also showed biallelic inactivation of TP53 and CDKN2A, findings which also appear common in visceral cases. One patient died of disease at 18 months after diagnosis, consistent with the aggressive nature of this tumor. Our results expand the anatomic spectrum of SD-UMN, adding this entity to an already challenging differential diagnosis that includes melanoma, squamous cell carcinoma, Merkel cell carcinoma, epithelioid sarcoma, and others. Given the potentially aggressive nature of SD-UMN, the timely and accurate diagnosis of this entity may have implications for prognosis and therapy.
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- 2022
12. Molecular and immunophenotypic characterization of SMARCB1 (INI1) - deficient intrathoracic Neoplasms
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Martina Haberecker, Marco Matteo Bühler, Alicia Pliego Mendieta, Roman Guggenberger, Fabian Arnold, Eva Markert, Markus Rechsteiner, Martin Zoche, Christian Britschgi, Chantal Pauli, University of Zurich, and Pauli, Chantal
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DNA Helicases ,Nuclear Proteins ,Sarcoma ,610 Medicine & health ,SMARCB1 Protein ,Chromatin Assembly and Disassembly ,Immunohistochemistry ,Pathology and Forensic Medicine ,2734 Pathology and Forensic Medicine ,10049 Institute of Pathology and Molecular Pathology ,10032 Clinic for Oncology and Hematology ,Biomarkers, Tumor ,Humans ,Retrospective Studies ,Transcription Factors - Abstract
The switch/sucrose-non-fermenting (SWI/SNF) complex is an ATP-dependent chromatin remodeling complex that plays important roles in DNA repair, transcription and cell differentiation. This complex consists of multiple subunits and is of particular interest in thoracic malignancies due to frequent subunit alteration of SMARCA4 (BRG1). Much less is known about SMARCB1 (INI1) deficient intrathoracic neoplasms, which are rare, often misclassified and understudied. In a retrospective analysis of 1479 intrathoracic malignant neoplasms using immunohistochemistry for INI1 (SMARCB1) on tissue micro arrays (TMA) and a search through our hospital sarcoma database, we identified in total nine intrathoracic, INI1 deficient cases (n = 9). We characterized these cases further by additional immunohistochemistry, broad targeted genomic analysis, methylation profiling and correlated them with clinical and radiological data. This showed that genomic SMARCB1 together with tumor suppressor alterations drive tumorigenesis in some of these cases, rather than epigenetic changes such as DNA methylation. A proper diagnostic classification, however, remains challenging. Intrathoracic tumors with loss or alteration of SMARCB1 (INI1) are highly aggressive and remain often underdiagnosed due to their rarity, which leads to false diagnostic interpretations. A better understanding of these tumors and proper diagnosis is important for better patient care as clinical trials and more targeted therapeutic options are emerging.
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- 2022
13. CDCA7 promotes TGF‐β‐induced epithelial–mesenchymal transition via transcriptionally regulating Smad4/Smad7 in ESCC
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Hongyi Li, Shaojie Wang, Xiubo Li, Yongjia Weng, Dinghe Guo, Pengzhou Kong, Caixia Cheng, Yanqiang Wang, Ling Zhang, Xiaolong Cheng, and Yongping Cui
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Cancer Research ,Epithelial-Mesenchymal Transition ,Esophageal Neoplasms ,Nuclear Proteins ,General Medicine ,Cadherins ,Smad7 Protein ,Gene Expression Regulation, Neoplastic ,Oncology ,Transforming Growth Factor beta ,Cell Movement ,Cell Line, Tumor ,Humans ,Vimentin ,Esophageal Squamous Cell Carcinoma ,Smad4 Protein - Abstract
Cell division cycle associated 7 (CDCA7) is a copy number amplification gene that contributes to the metastasis and invasion of tumors, including esophageal squamous cell carcinoma (ESCC). This present study aimed at clarifying whether high expression of CDCA7 promotes the metastasis and invasion of ESCC cell lines and exploring the underlying mechanisms implicated in epithelial-mesenchymal transition (EMT) of ESCC. The role of CDCA7 in the regulation of ESCC metastasis and invasion was evaluated using ESCC cell lines. Expression of EMT-related markers including E-cadherin, N-cadherin, Vimentin, Snail, and Slug, transforming growth factor β (TGF-β) signaling pathway including Smad2/3, p-Smad2/3, Smad4, and Smad7 were detected in CDCA7 knockdown and overexpressed cell lines. Dual-luciferase reporter assay and rescue assay were used to explore the underlying mechanisms that CDCA7 contributed to the metastasis and invasion of ESCC. High CDCA7 expression significantly promoted the metastasis and invasion of ESCC cell lines both in vivo and in vitro. Additionally, the expression of CDCA7 positively correlated with the expression of N-cadherin, Vimentin, Snail, Slug, TGF-β signaling pathway and negatively correlated with the expression of E-cadherin. Furthermore, CDCA7 transcriptionally regulated the expression of Smad4 and Smad7. Knockdown of CDCA7 inhibited the TGF-β signaling pathway and therefore inhibited EMT. Our data indicated that CDCA7 was heavily involved in EMT by regulating the expression of Smad4 and Smad7 in TGF-β signaling pathway. CDCA7 might be a new therapeutic target in the suppression of metastasis and invasion of ESCC.
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- 2022
14. BAF Complex Maintains Glioma Stem Cells in Pediatric H3K27M Glioma
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Eshini Panditharatna, Joana G. Marques, Tingjian Wang, Maria C. Trissal, Ilon Liu, Li Jiang, Alexander Beck, Andrew Groves, Neekesh V. Dharia, Deyao Li, Samantha E. Hoffman, Guillaume Kugener, McKenzie L. Shaw, Hafsa M. Mire, Olivia A. Hack, Joshua M. Dempster, Caleb Lareau, Lingling Dai, Logan H. Sigua, Michael A. Quezada, Ann-Catherine J. Stanton, Meghan Wyatt, Zohra Kalani, Amy Goodale, Francisca Vazquez, Federica Piccioni, John G. Doench, David E. Root, Jamie N. Anastas, Kristen L. Jones, Amy Saur Conway, Sylwia Stopka, Michael S. Regan, Yu Liang, Hyuk-Soo Seo, Kijun Song, Puspalata Bashyal, William P. Jerome, Nathan D. Mathewson, Sirano Dhe-Paganon, Mario L. Suvà, Angel M. Carcaboso, Cinzia Lavarino, Jaume Mora, Quang-De Nguyen, Keith L. Ligon, Yang Shi, Sameer Agnihotri, Nathalie Y.R. Agar, Kimberly Stegmaier, Charles D. Stiles, Michelle Monje, Todd R. Golub, Jun Qi, and Mariella G. Filbin
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Mammals ,Epigenome ,Oncology ,Mutation ,Neoplastic Stem Cells ,DNA Helicases ,Animals ,Humans ,Nuclear Proteins ,Glioma ,Transcription Factors - Abstract
Diffuse midline gliomas are uniformly fatal pediatric central nervous system cancers that are refractory to standard-of-care therapeutic modalities. The primary genetic drivers are a set of recurrent amino acid substitutions in genes encoding histone H3 (H3K27M), which are currently undruggable. These H3K27M oncohistones perturb normal chromatin architecture, resulting in an aberrant epigenetic landscape. To interrogate for epigenetic dependencies, we performed a CRISPR screen and show that patient-derived H3K27M-glioma neurospheres are dependent on core components of the mammalian BAF (SWI/SNF) chromatin remodeling complex. The BAF complex maintains glioma stem cells in a cycling, oligodendrocyte precursor cell–like state, in which genetic perturbation of the BAF catalytic subunit SMARCA4 (BRG1), as well as pharmacologic suppression, opposes proliferation, promotes progression of differentiation along the astrocytic lineage, and improves overall survival of patient-derived xenograft models. In summary, we demonstrate that therapeutic inhibition of the BAF complex has translational potential for children with H3K27M gliomas. Significance: Epigenetic dysregulation is at the core of H3K27M-glioma tumorigenesis. Here, we identify the BRG1–BAF complex as a critical regulator of enhancer and transcription factor landscapes, which maintain H3K27M glioma in their progenitor state, precluding glial differentiation, and establish pharmacologic targeting of the BAF complex as a novel treatment strategy for pediatric H3K27M glioma.
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- 2022
15. NCAPH Stabilizes GEN1 in Chromatin to Resolve Ultra-Fine DNA Bridges and Maintain Chromosome Stability
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Yuna Youn, Jae Hyeong Kim, and Jin-Hyeok Hwang
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Chromosomal Instability ,Humans ,Nuclear Proteins ,Cell Cycle Proteins ,DNA ,Cell Biology ,General Medicine ,Molecular Biology ,Chromatin ,Chromosomes - Abstract
Repairing damaged DNA and removing all physical connections between sister chromosomes is important to ensure proper chromosomal segregation by contributing to chromosomal stability. Here, we show that the depletion of non-SMC condensin I complex subunit H (NCAPH) exacerbates chromosome segregation errors and cytokinesis failure owing to sister-chromatid intertwinement, which is distinct from the ultra-fine DNA bridges induced by DNA inter-strand crosslinks (DNA-ICLs). Importantly, we identified an interaction between NCAPH and GEN1 in the chromatin involving binding at the N-terminus of NCAPH. DNA-ICL activation, using ICL-inducing agents, increased the expression and interaction between NCAPH and GEN1 in the soluble nuclear and chromatin, indicating that the NCAPH-GEN1 interaction participates in repairing DNA damage. Moreover, NCAPH stabilizes GEN1 within chromatin at the G2/M-phase and is associated with DNA-ICL-induced damage repair. Therefore, NCAPH resolves DNA-ICL-induced ultra-fine DNA bridges by stabilizing GEN1 and ensures proper chromosome separation and chromosome structural stability.
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- 2022
16. Mapping Ligand Interactions of Bromodomains BRD4 and ATAD2 with FragLites and PepLites─Halogenated Probes of Druglike and Peptide-like Molecular Interactions
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Gemma Davison, Mathew P. Martin, Shannon Turberville, Selma Dormen, Richard Heath, Amy B. Heptinstall, Marie Lawson, Duncan C. Miller, Yi Min Ng, James N. Sanderson, Ian Hope, Daniel J. Wood, Céline Cano, Jane A. Endicott, Ian R. Hardcastle, Martin E. M. Noble, and Michael J. Waring
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Binding Sites ,Protein Domains ,Drug Discovery ,Nuclear Proteins ,Molecular Medicine ,Cell Cycle Proteins ,Ligands ,Crystallography, X-Ray ,Peptides ,Transcription Factors ,Protein Binding - Abstract
The development of ligands for biological targets is critically dependent on the identification of sites on proteins that bind molecules with high affinity. A set of compounds, called FragLites, can identify such sites, along with the interactions required to gain affinity, by X-ray crystallography. We demonstrate the utility of FragLites in mapping the binding sites of bromodomain proteins BRD4 and ATAD2 and demonstrate that FragLite mapping is comparable to a full fragment screen in identifying ligand binding sites and key interactions. We extend the FragLite set with analogous compounds derived from amino acids (termed PepLites) that mimic the interactions of peptides. The output of the FragLite maps is shown to enable the development of ligands with leadlike potency. This work establishes the use of FragLite and PepLite screening at an early stage in ligand discovery allowing the rapid assessment of tractability of protein targets and informing downstream hit-finding.
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- 2022
17. Identification of a novel GR-ARID1a-P53BP1 protein complex involved in DNA damage repair and cell cycle regulation
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Stubbs, Felicity E, Flynn, Benjamin P, Rivers, Caroline A, Birnie, Matthew T, Herman, Andrew, Swinstead, Erin E, Baek, Songjoon, Fang, Hai, Temple, Jillian, Carroll, Jason S, Hager, Gordon L, Lightman, Stafford L, Conway-Campbell, Becky L, Flynn, Benjamin P [0000-0002-1871-3039], Rivers, Caroline A [0000-0002-2993-3723], Fang, Hai [0000-0003-3961-8572], Carroll, Jason S [0000-0003-3643-0080], Lightman, Stafford L [0000-0002-8546-9646], Conway-Campbell, Becky L [0000-0002-8494-7431], and Apollo - University of Cambridge Repository
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Cancer Research ,DNA Repair ,Cell Cycle ,Nuclear Proteins ,Cell Cycle Checkpoints ,Chromatin ,DNA-Binding Proteins ,Receptors, Glucocorticoid ,Cell Line, Tumor ,Genetics ,Humans ,Tumor Suppressor p53-Binding Protein 1 ,Molecular Biology ,DNA Damage ,Histone Acetyltransferases ,Transcription Factors - Abstract
Funder: RCUK | Medical Research Council (MRC); doi: https://doi.org/10.13039/501100000265, Funder: Wellcome Trust (Wellcome); doi: https://doi.org/10.13039/100004440, Funder: Office of the Assistant Secretary of Defense for Health Affairs. Breast Cancer Research Program. Award No. W81XWH-17-1-0067, ARID1a (BAF250), a component of human SWI/SNF chromatin remodeling complexes, is frequently mutated across numerous cancers, and its loss of function has been putatively linked to glucocorticoid resistance. Here, we interrogate the impact of siRNA knockdown of ARID1a compared to a functional interference approach in the HeLa human cervical cancer cell line. We report that ARID1a knockdown resulted in a significant global decrease in chromatin accessibility in ATAC-Seq analysis, as well as affecting a subset of genome-wide GR binding sites determined by analyzing GR ChIP-Seq data. Interestingly, the specific effects on gene expression were limited to a relatively small subset of glucocorticoid-regulated genes, notably those involved in cell cycle regulation and DNA repair. The vast majority of glucocorticoid-regulated genes were largely unaffected by ARID1a knockdown or functional interference, consistent with a more specific role for ARID1a in glucocorticoid function than previously speculated. Using liquid chromatography-mass spectrometry, we have identified a chromatin-associated protein complex comprising GR, ARID1a, and several DNA damage repair proteins including P53 binding protein 1 (P53BP1), Poly(ADP-Ribose) Polymerase 1 (PARP1), DNA damage-binding protein 1 (DDB1), DNA mismatch repair protein MSH6 and splicing factor proline and glutamine-rich protein (SFPQ), as well as the histone acetyltransferase KAT7, an epigenetic regulator of steroid-dependent transcription, DNA damage repair and cell cycle regulation. Not only was this protein complex ablated with both ARID1a knockdown and functional interference, but spontaneously arising DNA damage was also found to accumulate in a manner consistent with impaired DNA damage repair mechanisms. Recovery from dexamethasone-dependent cell cycle arrest was also significantly impaired. Taken together, our data demonstrate that although glucocorticoids can still promote cell cycle arrest in the absence of ARID1a, the purpose of this arrest to allow time for DNA damage repair is hindered.
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- 2022
18. How retinoic acid and arsenic transformed acute promyelocytic leukemia therapy
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Victoria Korsos and Wilson H Miller Jr
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Receptors, Retinoic Acid ,Retinoic Acid Receptor alpha ,Tumor Suppressor Proteins ,Nuclear Proteins ,Tretinoin ,Promyelocytic Leukemia Protein ,Translocation, Genetic ,Arsenic ,Endocrinology ,Arsenic Trioxide ,Leukemia, Promyelocytic, Acute ,Humans ,Molecular Biology ,In Situ Hybridization, Fluorescence ,Transcription Factors - Abstract
Acute promyelocytic leukemia (APL) is associated with severe coagulopathy leading to rapid morbidity and mortality if left untreated. The definitive diagnosis of APL is made by identifying a balanced reciprocal translocation between chromosomes 15 and 17. This t(15;17) results in a fusion transcript of promyelocytic leukemia (PML) and retinoic acid receptor alpha (RARA) genes and the expression of a functional PML/RARA protein. Detection of a fused PML/RARA genomic DNA sequence using fluorescence in situ hybridization (FISH) or by detection of the PML/RARA fusion transcript via reverse transcriptase polymerase chain reaction (RT-PCR) has revolutionized the diagnosis and monitoring of APL. Once confirmed, APL is cured in over 90% of cases, making it the most curable subtype of acute leukemia today. Patients with low-risk APL are successfully treated using a chemotherapy-free combination of all-trans retinoic acid and arsenic trioxide (ATO). In this review, we explore the work that has gone into the modern-day diagnosis and highly successful treatment of this once devastating leukemia.
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- 2022
19. BET inhibition triggers antitumor immunity by enhancing MHC class I expression in head and neck squamous cell carcinoma
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Ming Zhang, Ganping Wang, Zhikun Ma, Gan Xiong, Wenjin Wang, Zhengxian Huang, Yuehan Wan, Xiuyun Xu, Rosalie G. Hoyle, Chen Yi, Jinsong Hou, Xiqiang Liu, Demeng Chen, Jiong Li, and Cheng Wang
- Subjects
Pharmacology ,Squamous Cell Carcinoma of Head and Neck ,Histocompatibility Antigens Class I ,Nuclear Proteins ,Cell Cycle Proteins ,CD8-Positive T-Lymphocytes ,Head and Neck Neoplasms ,Cell Line, Tumor ,Drug Discovery ,Genetics ,Humans ,Molecular Medicine ,Molecular Biology ,Transcription Factors - Abstract
BET inhibition has been shown to have a promising antitumor effect in multiple tumors. However, the impact of BET inhibition on antitumor immunity was still not well documented in HNSCC. In this study, we aim to assess the functional role of BET inhibition in antitumor immunity and clarify its mechanism. We show that BRD4 is highly expressed in HNSCC and inversely correlated with the infiltration of CD8sup+/supT cells. BET inhibition potentiates CD8sup+/supT cell-based antitumor immunity in vitro and in vivo. Mechanistically, BRD4 acts as a transcriptional suppressor and represses the expression of MHC class I molecules by recruiting G9a. Pharmacological inhibition or genetic depletion of BRD4 potently increases the expression of MHC class I molecules in the absence and presence of IFN-γ. Moreover, compared to PD-1 blocking antibody treatment or JQ1 treatment individually, the combination of BET inhibition with anti-PD-1 antibody treatment significantly enhances the antitumor response in HNSCC. Taken together, our data unveil a novel mechanism by which BET inhibition potentiates antitumor immunity via promoting the expression of MHC class I molecules and provides a rationale for the combination of ICBs with BET inhibitors for HNSCC treatment.
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- 2022
20. Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells
- Author
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Shipeng Chen, Lian Guan, Xu Zhao, Jing Yang, Longqing Chen, Mengmeng Guo, Juanjuan Zhao, Chao Chen, Ya Zhou, Yong Han, and Lin Xu
- Subjects
Lung Neoplasms ,General Veterinary ,Thyroid Nuclear Factor 1 ,Thyroid Gland ,Nuclear Proteins ,General Medicine ,General Biochemistry, Genetics and Molecular Biology ,Mice ,MicroRNAs ,Carcinoma, Non-Small-Cell Lung ,Animals ,Humans ,General Pharmacology, Toxicology and Pharmaceutics ,Research Article ,Transcription Factors - Abstract
Targeted gene therapy has become a promising approach for lung cancer treatment. In our previous work, we reported that the targeted expression of microRNA-7 (miR-7) operated by thyroid transcription factor-1 (TTF-1) promoter inhibited the growth of human lung cancer cells in vitro and in vivo; however, the intervention efficiency needed to be further improved. In this study, we identified the core promoter of TTF-1 (from -1299 bp to -871 bp) by 5' deletion assay and screened out the putative transcription factors nuclear factor-1 (NF-1) and activator protein-1 (AP-1). Further analysis revealed that the expression level of NF-1, but not AP-1, was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer (NSCLC) cells. Moreover, the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter. Of note, we optimized four distinct sequences to form additional NF-1-binding sites (TGGCA) in the sequence of TTF-1 core promoter (termed as (opt) TTF-1 promoter), and verified the binding efficiency of NF-1 on the (opt) TTF-1 promoter by electrophoretic mobility shift assay (EMSA). As expected, the (opt) TTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro, accompanied by a reduced transduction of NADH dehydrogenase (ubiquinone) 1α subcomplex 4 (NDUFA4)/protein kinase B (Akt) pathway. Consistently, (opt) TTF-1 promoter-driven miR-7 expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC. Finally, no significant changes were detected in the biological indicators or the histology of some important tissues and organs, including heart, liver, and spleen. On the whole, our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells, providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.
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- 2022
21. Strain stiffening of Ndc80 complexes attached to microtubule plus ends
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Felix Schwietert, Vladimir A. Volkov, Pim J. Huis in ’t Veld, Marileen Dogterom, Andrea Musacchio, and Jan Kierfeld
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Chromosome Segregation ,Biophysics ,Nuclear Proteins ,Spindle Apparatus ,Kinetochores ,Microtubules - Abstract
In the mitotic spindle, microtubules attach to chromosomes via kinetochores. The microtubule-binding Ndc80 complex is an integral part of kinetochores, and is essential for kinetochores to attach to microtubules and to transmit forces from dynamic microtubule ends to the chromosomes. The Ndc80 complex has a rod-like appearance with globular domains at its ends that are separated by a long coiled coil. Its mechanical properties are considered important for the dynamic interaction between kinetochores and microtubules. Here, we present a novel method that allows us to time-trace the effective stiffness of Ndc80 complexes following shortening microtubule ends against applied force in optical trap experiments. Applying this method to wild type Ndc80 and three variants (CH-domains mutated or Hec1-tail unphosphorylated, phosphorylated, or truncated), we reveal that each variant exhibits strain stiffening, i.e., the effective stiffness increases under tension that is built up by a depolymerizing microtubule. The strain stiffening relation is roughly linear and independent of the state of the microtubule. We introduce structure-based models, which show that the strain stiffening can be traced back to the specific architecture of the Ndc80 complex with a characteristic flexible kink, to thermal fluctuations of the microtubule, and to the bending elasticity of flaring protofilaments, which exert force to move the Ndc80 complexes. Our model accounts for changes in the amount of load-bearing attachments at various force levels and reproduces the roughly linear strain stiffening behavior, highlighting the importance of force-dependent binding affinity.SIGNIFICANCEBy time-tracing the stiffness of microtubule end-tracking Ndc80 complexes in optical trap experiments, we detect strain stiffening, and, thereby, provide new insights into the elastic properties of the Ndc80 complex. The strain stiffening is robust against mutations in the Ndc80 complex. We relate strain stiffening to the structure of the Ndc80 complex by means of a simple polymer model, to thermal fluctuations of the microtubule, and to the flexibility of force-generating flaring protofilaments at the tip of the microtubule. Since Ndc80 complexes play a major role for transmitting force from microtubule ends to the kinetochore, their elastic properties are of great interest for a deeper understanding of chromosome dynamics in the mitotic spindle.
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- 2022
22. The diagnostic and prognostic utility of incorporating DAXX, ATRX, and alternative lengthening of telomeres to the evaluation of pancreatic neuroendocrine tumors
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Christopher M. Heaphy and Aatur D. Singhi
- Subjects
Pancreatic Neoplasms ,Neuroendocrine Tumors ,X-linked Nuclear Protein ,Biomarkers, Tumor ,Humans ,Nuclear Proteins ,Telomere ,Prognosis ,Co-Repressor Proteins ,In Situ Hybridization, Fluorescence ,Adaptor Proteins, Signal Transducing ,Molecular Chaperones ,Pathology and Forensic Medicine - Abstract
Pancreatic neuroendocrine tumors (PanNETs) are a heterogeneous group of neoplasms with increasing incidence and an ill-defined pathobiology. Although many PanNETs are indolent and remain stable for years, a subset may behave aggressively and metastasize widely. Thus, the increasing and frequent detection of PanNETs presents a treatment dilemma. Current prognostic systems are susceptible to interpretation errors, sampling issues, and do not accurately reflect the clinical behavior of these neoplasms. Hence, additional biomarkers are needed to improve the prognostic stratification of patients diagnosed with a PanNET. Recent studies have identified alterations in death domain-associated protein 6 (DAXX) and alpha-thalassemia/mental retardation X-linked (ATRX), as well as alternative lengthening of telomeres (ALT), as promising prognostic biomarkers. This review summarizes the identification, clinical utility, and specific nuances in testing for DAXX/ATRX by immunohistochemistry and ALT by telomere-specific fluorescence in situ hybridization in PanNETs. Furthermore, a discussion on diagnostic indications for DAXX, ATRX, and ALT status is provided to include the distinction between PanNETs and pancreatic neuroendocrine carcinomas (PanNECs), and determining pancreatic origin for metastatic neuroendocrine tumors in the setting of an unknown primary.
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- 2022
23. A Simulation Study of the Effect of Naturally Occurring Point Mutations on the SRY-DNA Complex
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Nasou, Angeliki-Georgia, Pantatosaki, Evangelia, and Papadopoulos, George K.
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DNA-Binding Proteins ,Materials Chemistry ,Humans ,Point Mutation ,Nuclear Proteins ,Amino Acid Sequence ,DNA ,Physical and Theoretical Chemistry ,Sex-Determining Region Y Protein ,Surfaces, Coatings and Films - Abstract
Molecular dynamics (MD) simulations were conducted in order to investigate the effect of the naturally occurring point mutations of the transcription factor (TF) sex-determining region Y (SRY) on the structure and dynamics of the SRY-DNA complex. The normal SRY, along with the two mutants I13T and G40R, comprising point mutations on the SRY chain, which have been clinically identified in patients with sex developmental disorders, were modeled as DNA complexes. Our modeling work aims at elucidating atomic-level structural determinants of the aberrant SRY-DNA complexation by means of μs-long MD. The results suggest that the observed disorders brought about by the G40R-DNA and I13T-DNA may arise predominantly from the destabilization of the complex being in accord with
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- 2022
24. Modeling Study of the Dynamics of Kinesin-14 Molecular Motors
- Author
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Ping Xie
- Subjects
Saccharomyces cerevisiae Proteins ,Microtubule Proteins ,Materials Chemistry ,Animals ,Humans ,Kinesins ,Nuclear Proteins ,Drosophila ,Saccharomyces cerevisiae ,Physical and Theoretical Chemistry ,Kinetochores ,Microtubules ,Surfaces, Coatings and Films - Abstract
Kinesin-14s constitute a subfamily of the large superfamily of adenosine triphosphate-dependent microtubule-based motor proteins. Kinesin-14s have the motor domain at the C-terminal end of the peptide, playing key roles during spindle assembly and maintenance. Some of them are nonprocessive motors, whereas others can move processively on microtubules. Here, we take budding yeast Cik1-Kar3 and human HSET as examples to study theoretically the dynamics of the processive kinesin-14 motor moving on the single microtubule under load, the dynamics of the motor coupled with an Ndc80 protein moving on the single microtubule, the dynamics of the motor moving in microtubule arrays, and so on. The dynamics of the nonprocessive
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- 2022
25. The H3K9me2‐binding protein AGDP3 limits DNA methylation and transcriptional gene silencing in Arabidopsis
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Xuelin Zhou, Mengwei Wei, Wenfeng Nie, Yue Xi, Li Peng, Qijie Zheng, Kai Tang, Viswanathan Satheesh, Yuhua Wang, Jinyan Luo, Xuan Du, Rui Liu, Zhenlin Yang, Honggui La, Yingli Zhong, Yu Yang, Jian‐Kang Zhu, Jiamu Du, and Mingguang Lei
- Subjects
Arabidopsis Proteins ,Proto-Oncogene Proteins ,Arabidopsis ,Nuclear Proteins ,Gene Silencing ,DNA ,Plant Science ,DNA Methylation ,Protein-Tyrosine Kinases ,Carrier Proteins ,Biochemistry ,General Biochemistry, Genetics and Molecular Biology - Abstract
DNA methylation, a conserved epigenetic mark, is critical for tuning temporal and spatial gene expression. The Arabidopsis thaliana DNA glycosylase/lyase REPRESSOR OF SILENCING 1 (ROS1) initiates active DNA demethylation and is required to prevent DNA hypermethylation at thousands of genomic loci. However, how ROS1 is recruited to specific loci is not well understood. Here, we report the discovery of Arabidopsis AGENET Domain Containing Protein 3 (AGDP3) as a cellular factor that is required to prevent gene silencing and DNA hypermethylation. AGDP3 binds to H3K9me2 marks in its target DNA via its AGD12 cassette. Analysis of the crystal structure of the AGD12 cassette of AGDP3 in complex with an H3K9me2 peptide revealed that dimethylated H3K9 and unmodified H3K4 are specifically anchored into two different surface pockets. A histidine residue located in the methyllysine binding aromatic cage provides AGDP3 with pH-dependent H3K9me2 binding capacity. Our results uncover a molecular mechanism for the regulation of DNA demethylation by the gene silencing mark H3K9me2.
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- 2022
26. Bioinformatics Analysis of Common Genetic and Molecular Traits and Association of Portal Hypertension with Pulmonary Hypertension
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MingYu Chen and YouPeng Chen
- Subjects
Inflammation ,Article Subject ,Hypertension, Pulmonary ,Biomedical Engineering ,Computational Biology ,Endothelial Cells ,Nuclear Proteins ,Health Informatics ,MicroRNAs ,GTP-Binding Proteins ,Hypertension, Portal ,Humans ,Surgery ,Tumor Suppressor Protein p53 ,Biotechnology - Abstract
Portal hypertension (PH) is an important cause of pulmonary arterial hypertension(PAH), but its mechanism is still unclear. We used genetic data analysis to explore the shared genes and molecular mechanisms of PH and PAH. We downloaded the PH and PAH data from the GEO database, and used the weighted gene coexpression network analysis method (WGCNA) to analyze the coexpression modules of idiopathic noncirrhotic portal hypertension (INCPH) and cirrhotic portal hypertension (CPH) and pulmonary hypertension, respectively. Enrichment analysis was performed on the common genes, and differential gene expressions (DEGs) were used for verification. The target genes of INCPH and PAH were obtained by string and cytoscape software, and the miRNAs of target genes were predicted by miRwalk, miRDB, and TargetScan and their biological functions were analyzed; finally, we used PanglaoDB to predict the expression of target genes in cells. In WGCNA, gene modules significantly related to PAH, CPH, and INCPH were identified, and enrichment function analysis showed that the common pathway of PAH and CPH were “P53 signaling pathway,” “synthesis of neutral lipids”; PAH and INCPH are “terminal,” “Maintenance Regulation of Granules,” and “Toxin Transport.” DEGs confirmed the results of WGCNA; the common miRNA functions of PAH and cirrhosis were enriched for “P53 signaling pathway,” “TGF-β signaling pathway,” “TNF signaling pathway,” and “fatty acid metabolism,” and the miRNAs-mRNAs network suggested that hsa-miR-22a-3p regulates MDM2 and hsa-miR-34a-5p regulates PRDX4; the target genes of PAH and INCPH are EIF5B, HSPA4, GNL3, RARS, UTP20, HNRNPA2B1, HSP90B1, METAP2, NARS, SACM1L, and their target miRNA function enrichment showed EIF5B, HNRNPA2B1, HSP90B1, METAP2, NARS, SACM1L, and HSPA4 are associated with telomeres and inflammation, panglaoDB showed that target genes are located in endothelial cells, smooth muscle cells, etc. In conclusion, the mechanism of pulmonary hypertension induced by portal hypertension may be related to telomere dysfunction and P53 overactivation, and lipid metabolism and intestinal inflammation are also involved in this process.
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- 2022
27. Comprehensive Analysis of the Oncogenic Role of Targeting Protein for Xklp2 (TPX2) in Human Malignancies
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Ting Shao, Xiling Jiang, Guochang Bao, Chunsheng Li, and Changgang Guo
- Subjects
Carcinoma, Transitional Cell ,Carcinoma, Hepatocellular ,Article Subject ,Liver Neoplasms ,Biochemistry (medical) ,Clinical Biochemistry ,Nuclear Proteins ,Cell Cycle Proteins ,General Medicine ,Urinary Bladder Neoplasms ,Biomarkers, Tumor ,Tumor Microenvironment ,Genetics ,Humans ,Microsatellite Instability ,Microtubule-Associated Proteins ,Molecular Biology - Abstract
Mitosis and spindle assembly require the microtubule-associated protein Xenopus kinesin-like protein 2 (TPX2). Although TPX2 is highly expressed in several malignant tumor forms, little is known about its role in cancer. In this study, we performed the gene set enrichment analysis of TPX2 in 33 types of cancers and an extensive pan-cancer bioinformatic analysis using prognosis, tumor mutational burdens, microsatellite instability, tumor microenvironment, and immune cell infiltration data. According to the differential expression study, TPX2 was found to be overexpressed across all studied cancer types. Based on the survival analysis, increased TPX2 expression was associated with a poor prognosis for most cancers. The TPX2 expression level was confirmed to correlate with the clinical stage, microsatellite instability, and tumor mutational burden across all cancer types. Furthermore, TPX2 expression has been linked to tumor microenvironments and immune cell infiltration, particularly in bladder urothelial carcinoma, liver hepatocellular carcinoma, lung adenocarcinoma, stomach adenocarcinoma, and uterine corpus endometrial carcinoma. Finally, the gene set enrichment analysis implicated TPX2 in the regulation of aminoacyl tRNA biosynthesis, which is the most important tumor cell cycle signaling pathway. This comprehensive pan-cancer analysis shows that TPX2 is a prognostic molecular biomarker for most cancers and suggests its potential as an effective therapeutic target for the treatment of these diseases.
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- 2022
28. Prognostic impact of NPM1 and FLT3 mutations in patients with AML in first remission treated with oral azacitidine
- Author
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Hartmut Döhner, Andrew H. Wei, Gail J. Roboz, Pau Montesinos, Felicitas R. Thol, Farhad Ravandi, Hervé Dombret, Kimmo Porkka, Irwindeep Sandhu, Barry Skikne, Wendy L. See, Manuel Ugidos, Alberto Risueño, Esther T. Chan, Anjan Thakurta, C.L. Beach, and Daniel Lopes de Menezes
- Subjects
Neoplasm, Residual ,Remission Induction ,Immunology ,Nuclear Proteins ,Cell Biology ,Hematology ,Protein-Tyrosine Kinases ,Prognosis ,Biochemistry ,Leukemia, Myeloid, Acute ,fms-Like Tyrosine Kinase 3 ,Recurrence ,Mutation ,Azacitidine ,Humans ,Nucleophosmin - Abstract
The randomized, placebo-controlled, phase 3 QUAZAR AML-001 trial (ClinicalTrials.gov identifier: NCT01757535) evaluated oral azacitidine (Oral-AZA) in patients with acute myeloid leukemia (AML) in first remission after intensive chemotherapy (IC) who were not candidates for hematopoietic stem cell transplantation. Eligible patients were randomized 1:1 to Oral-AZA 300 mg or placebo for 14 days per 28-day cycle. We evaluated relapse-free survival (RFS) and overall survival (OS) in patient subgroups defined by NPM1 and FLT3 mutational status at AML diagnosis and whether survival outcomes in these subgroups were influenced by presence of post-IC measurable residual disease (MRD). Gene mutations at diagnosis were collected from patient case report forms; MRD was determined centrally by multiparameter flow cytometry. Overall, 469 of 472 randomized patients (99.4%) had available mutational data; 137 patients (29.2%) had NPM1 mutations (NPM1mut), 66 patients (14.1%) had FLT3 mutations (FLT3mut; with internal tandem duplications [ITD], tyrosine kinase domain mutations [TKDmut], or both), and 30 patients (6.4%) had NPM1mut and FLT3-ITD at diagnosis. Among patients with NPM1mut, OS and RFS were improved with Oral-AZA by 37% (hazard ratio [HR], 0.63; 95% confidence interval [CI], 0.41-0.98) and 45% (HR, 0.55; 95% CI, 0.35-0.84), respectively, vs placebo. Median OS was improved numerically with Oral-AZA among patients with NPM1mut whether without MRD (48.6 months vs 31.4 months with placebo) or with MRD (46.1 months vs 10.0 months with placebo) post-IC. Among patients with FLT3mut, Oral-AZA improved OS and RFS by 37% (HR, 0.63; 95% CI, 0.35-1.12) and 49% (HR, 0.51; 95% CI, 0.27-0.95), respectively, vs placebo. Median OS with Oral-AZA vs placebo was 28.2 months vs 16.2 months, respectively, for patients with FLT3mut and without MRD and 24.0 months vs 8.0 months for patients with FLT3mut and MRD. In multivariate analyses, Oral-AZA significantly improved survival independent of NPM1 or FLT3 mutational status, cytogenetic risk, or post-IC MRD status.
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- 2022
29. <scp>RNA</scp> ‐sequencing analysis reveals the potential molecular mechanism of <scp>RAD54B</scp> in the proliferation of inflamed human dental pulp cells
- Author
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Pei Huang, Fushi Wang, Xinhuan Wang, Xiujiao Meng, Weiwei Qiao, and Liuyan Meng
- Subjects
DNA Helicases ,Humans ,RNA ,Nuclear Proteins ,Tumor Suppressor Protein p53 ,General Dentistry ,Dental Pulp ,Cell Proliferation - Abstract
To investigate the role of RAD54B in the proliferation of inflamed human dental pulp cells (hDPCs) induced by lipopolysaccharide (LPS).Normal, carious and pulpitic human dental pulp tissues were collected. Total RNA was subjected to RNA-sequencing (seq) and gene expression profiles were studied by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Differentially expressed genes (DEGs) in homologous recombination repair (HRR) were validated with qRT-PCR. The expression of RAD54B and TNF-α in human dental pulp tissues was detected using immunohistochemistry. HDPCs were cultured and RAD54B level in hDPCs was detected after LPS stimulation using western blot. CCK-8 was used to investigate the proliferation of hDPCs transfected with negative control (Nc) small interfering RNA (siRNA), RAD54B siRNA, P53 siRNA or both siRNAs with or without LPS stimulation. Flow cytometry was used to detect the cell cycle distribution, and western blot and immunofluorescence were used to analyse the expression of RAD54B, P53 and P21 under the above treatments. One-way and two-way anova followed by least significant difference posttest were used for statistical analysis.RNA-seq results identified DEGs amongst the three groups. KEGG pathway analysis revealed enrichment of DEGs in the replication and repair pathway. HRR and non-homologous end joining (NHEJ) components were further verified and qRT-PCR results were basically consistent with the sequencing data. RAD54B, an HRR accessory factor highly expressed in carious and pulpitic tissues as compared to that in normal pulps, was chosen as our gene of interest. High RAD54B expression was confirmed in inflamed human dental pulp tissues and LPS-stimulated hDPCs. Upon RAD54B knockdown, P53 and P21 expressions in hDPCs were upregulated whereas the proliferation was significantly downregulated, accompanied by increased G2/M phase arrest. After inhibiting P53 expression in RAD54B-knockdown hDPCs, P21 expression and cell proliferation were reversed.Gene expression profiles of normal, carious and pulpitic human dental pulp tissues were revealed. HRR components were elucidated to function in dental pulp inflammation. Amongst the DEGs in HRR, RAD54B regulated the proliferation of inflamed hDPCs via P53/P21 signalling. This research deepens our understanding of dental pulp inflammation and provides new insight to clarify the underlying mechanisms.
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- 2022
30. <scp>Hsa‐microRNA</scp> ‐370‐3p targeting Snail and Twist1 suppresses <scp>IL</scp> ‐8/ <scp>STAT3</scp> ‐driven hepatocellular carcinoma metastasis
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Siqi Peng, Yutong Chen, Ting Li, Junjie Mao, Pengfei Yang, Baojia Zou, Lisi Luo, Weiyu Zhang, Wen Wang, Rongzhi Xie, Jian Li, and Linjuan Zeng
- Subjects
STAT3 Transcription Factor ,Cancer Research ,Carcinoma, Hepatocellular ,Epithelial-Mesenchymal Transition ,Interleukin-8 ,Liver Neoplasms ,Twist-Related Protein 1 ,Mice, Nude ,Nuclear Proteins ,General Medicine ,Gene Expression Regulation, Neoplastic ,Mice ,MicroRNAs ,Oncology ,Cell Movement ,Cell Line, Tumor ,Animals ,Humans ,RNA, Messenger ,Cell Proliferation - Abstract
The pro-inflammatory factor interleukin-8 (IL-8) is related to poor prognosis in hepatocellular carcinoma (HCC) patients. Interleukin-8 enhanced HCC invasion by upregulating Snail and Twist1, whether this modulation relies on microRNAs (miR) is unclear. In this study, hsa-miR-370-3p was screened as candidate miRNA targeting Snail and Twist1, and its expression was downregulated by IL-8. Luciferase assays and RNA electrophoretic mobility shift assays were used to evaluate the interaction between miR-370-3p and targeted mRNAs. Coimmunoprecipitation, luciferase, and ChIP assays were undertaken to investigate the mechanisms underlying IL-8-mediated modification of miR-370-3p. Gain- and loss-of-function studies, Transwell assays, and a xenograft nude mouse model were used to investigate pro- and antitumor activities. Interleukin-8 and miR-370-3p levels were analyzed for clinical relevance in HCC patients. Our results showed that HCC patients with high levels of IL-8 experienced more metastasis and shorter survival. Interleukin-8 induced epithelial-mesenchymal transition and promoted liver cancer cell migration, invasion, and metastasis both in vitro and in vivo. MicroRNA-370-3p interacted with its cognate mRNA within the 3'-UTR regions of Twist1 and Snail mRNA directly and specifically and attenuated IL-8 protumoral effects on liver cancer cells. Interleukin-8 negatively modulated miR-370-3p through signal transducer and activator of transcription 3 (STAT3) activation by recruiting histone deacetylase 1 (HDAC1) to miR-370-3p promoter. The STAT3 and HDAC antagonists inhibited liver cancer cell migration and invasion. Patients with high miR-370-3p and low IL-8 levels had longer overall survival. In conclusion, our study elucidated a novel axis IL-8/STAT3/miR-370-3p/Twist1 and Snail relying on HDAC1 recruitment, which showed both diagnostic and therapeutic potentials of miR-370-3p in HCC metastasis.
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- 2022
31. Bromodomain Interactions with Acetylated Histone 4 Peptides in the BRD4 Tandem Domain: Effects on Domain Dynamics and Internal Flexibility
- Author
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Sven Wernersson, Romel Bobby, Liz Flavell, Alexander G. Milbradt, Geoffrey A. Holdgate, Kevin J. Embrey, and Mikael Akke
- Subjects
Histones ,Binding Sites ,Nuclear Proteins ,Cell Cycle Proteins ,Peptides ,Biochemistry ,Transcription Factors - Abstract
The bromodomain and extra-terminal (BET) protein BRD4 regulates gene expression via recruitment of transcriptional regulatory complexes to acetylated chromatin. Like other BET proteins, BRD4 contains two bromodomains, BD1 and BD2, that can interact cooperatively with target proteins and designed ligands, with important implications for drug discovery. Here, we used nuclear magnetic resonance (NMR) spectroscopy to study the dynamics and interactions of the isolated bromodomains, as well as the tandem construct including both domains and the intervening linker, and investigated the effects of binding a tetra-acetylated peptide corresponding to the tail of histone 4. The peptide affinity is lower for both domains in the tandem construct than for the isolated domains. Using
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- 2022
32. APE1 assembles biomolecular condensates to promote the ATR–Chk1 DNA damage response in nucleolus
- Author
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Jia Li, Haichao Zhao, Anne McMahon, and Shan Yan
- Subjects
Biomolecular Condensates ,DNA-Binding Proteins ,RNA, Ribosomal ,Checkpoint Kinase 1 ,Genetics ,Nuclear Proteins ,Ataxia Telangiectasia Mutated Proteins ,DNA ,RNA, Small Interfering ,DNA Damage - Abstract
Multifunctional protein APE1/APEX1/HAP1/Ref-1 (designated as APE1) plays important roles in nuclease-mediated DNA repair and redox regulation in transcription. However, it is unclear how APE1 regulates the DNA damage response (DDR) pathways and influences genome integrity directly or indirectly. Here we show that siRNA-mediated APE1-knockdown or APE1 inhibitor treatment attenuates the ATR-Chk1 DDR under stress conditions in multiple immortalized cell lines. Congruently, APE1 overexpression (APE1-OE) activates the ATR DDR under unperturbed conditions, which is independent of APE1 nuclease and redox functions. Structural and functional analysis reveals a direct requirement of the extreme N-terminal 33 amino acids (NT33) within APE1 in the assembly of distinct biomolecular condensates in vitro and DNA/RNA-independent activation of the ATR DDR. Overexpressed APE1 co-localizes with nucleolar NPM1 and assembles biomolecular condensates in nucleoli in cancer but not non-malignant cells, which recruits ATR and its direct activator molecules TopBP1 and ETAA1. APE1 W119R mutant is deficient in nucleolar condensation and liquid-liquid phase separation and is incapable of activating nucleolar ATR DDR. Lastly, APE1-OE-induced nucleolar ATR DDR activation leads to compromised ribosomal RNA transcription and reduced cell viability. Taken together, we propose distinct mechanisms by which APE1 regulates ATR DDR pathways and functions in genome integrity maintenance.
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- 2022
33. Hybrid consensus and k-nearest neighbours (kNN) strategies to classify dual BRD4/PLK1 inhibitors
- Author
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H. Rezaie, M. Asadollahi-Baboli, and S.K. Hassaninejad-Darzi
- Subjects
Consensus ,Drug Discovery ,Humans ,Nuclear Proteins ,Quantitative Structure-Activity Relationship ,Reproducibility of Results ,Molecular Medicine ,Cell Cycle Proteins ,Bioengineering ,General Medicine ,Transcription Factors - Abstract
A novel decision-making procedure is proposed here for the first time to identify active/inactive and selective/non-selective dual inhibitors using consensus approaches and pools of k-nearest neighbours (kNN) classifications instead of individual models. Dual BRD4/PLK1 inhibition with adequate selectivity is a potential therapeutic strategy for targeting tumour cells in high-risk patients. We report the unique way to identify both active and selective dual BRD4/PLK1 inhibitors using consensus and kNN strategies together with two sources of receptor-based and ligand-based information which are the ranked binding energies of residues and important molecular features, respectively. The results of consensus approaches were compared with the results of individual kNN models. The chemical space similarity was measured using three different distance functions to increase the reliability. All activity and selectivity classification models were validated using cross-validation and y-randomization tests. The outcomes show that consensus approaches can increase the reliability and accuracy of active/inactive or selective/non-selective detections up to 90%. Consensus approaches also reached more balanced values of sensitivity and specificity compared to the individual kNN models because of the compensation in the integration of diverse sources of information.
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- 2022
34. Telomere biology disorders: time for moving towards the clinic?
- Author
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Luis F.Z. Batista, Inderjeet Dokal, and Roy Parker
- Subjects
Mutation ,Humans ,Nuclear Proteins ,RNA ,RNA-Binding Proteins ,Molecular Medicine ,Cell Cycle Proteins ,Telomere ,Biology ,Telomerase ,Molecular Biology ,Dyskeratosis Congenita - Abstract
Telomere biology disorders (TBDs) are a group of rare diseases caused by mutations that impair telomere maintenance. Mutations that cause reduced levels of TERC/hTR, the telomerase RNA component, are found in most TBD patients and include loss-of-function mutations in hTR itself, in hTR-binding proteins [NOP10, NHP2, NAF1, ZCCHC8, and dyskerin (DKC1)], and in proteins required for hTR processing (PARN). These patients show diverse clinical presentations that most commonly include bone marrow failure (BMF)/aplastic anemia (AA), pulmonary fibrosis, and liver cirrhosis. There are no curative therapies for TBD patients. An understanding of hTR biogenesis, maturation, and degradation has identified pathways and pharmacological agents targeting the poly(A) polymerase PAPD5, which adds 3'-oligoadenosine tails to hTR to promote hTR degradation, and TGS1, which modifies the 5'-cap structure of hTR to enhance degradation, as possible therapeutic approaches. Critical next steps will be clinical trials to establish the effectiveness and potential side effects of these compounds in TBD patients.
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- 2022
35. Structural basis for the recognition by 14-3-3 proteins of a conditional binding site within the oligomerization domain of human nucleophosmin
- Author
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Anna A. Kapitonova, Kristina V. Tugaeva, Larisa A. Varfolomeeva, Konstantin M. Boyko, Richard B. Cooley, and Nikolai N. Sluchanko
- Subjects
Phosphopeptides ,Binding Sites ,14-3-3 Proteins ,Biophysics ,Humans ,Nuclear Proteins ,Cell Biology ,Nucleophosmin ,Molecular Biology ,Biochemistry - Abstract
Nucleophosmin 1 (NPM1) is a multifunctional protein regulating ribosome biogenesis, centrosome duplication and chromatin remodeling. Being a major nucleolar protein, NPM1 can migrate to the nucleus and the cytoplasm, which is controlled by changes of NPM1 oligomerization and interaction with other cell factors. NPM1 forms a stable pentamer with its N-terminal structured domain, where two nuclear export signals and several phosphorylation sites reside. This domain undergoes dissociation and disordering upon Ser48 phosphorylation in the subunit interface. Recent studies indicated that Ser48 is important for NPM1 interaction with other proteins including 14-3-3, the well-known phosphoserine/phosphothreonine binders, but the structural basis for 14-3-3/NPM1 interaction remained unaddressed. By fusing human 14-3-3ζ with an NPM1 segment surrounding Ser48, which was phosphorylated inside Escherichia coli cells by co-expressed protein kinase A, here we obtained the desired protein/phosphopeptide complex and determined its crystal structure. While biochemical data indicated that the interaction is driven by Ser48 phosphorylation, the crystallographic 14-3-3/phosphopeptide interface reveals an NPM1 conformation distinctly different from that in the NPM1 pentamer. Given the canonical phosphopeptide-binding mode observed in our crystal structure, Ser48 emerges as a conditional binding site whose recognition by 14-3-3 proteins is enabled by NPM1 phosphorylation, disassembly and disordering under physiological circumstances.
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- 2022
36. JQ1 attenuates neuroinflammation by inhibiting the inflammasome-dependent canonical pyroptosis pathway in SAE
- Author
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Xiaolin Zhong, Zuyao Chen, Yajuan Wang, Mingli Mao, Yingcheng Deng, Mengmeng Shi, Yang Xu, Ling Chen, and Wenyu Cao
- Subjects
Lipopolysaccharides ,Inflammasomes ,General Neuroscience ,NF-kappa B ,Nuclear Proteins ,Azepines ,Triazoles ,Mice ,Occludin ,Sepsis ,Sepsis-Associated Encephalopathy ,Neuroinflammatory Diseases ,Pyroptosis ,Animals ,Transcription Factors - Abstract
Sepsis-associated encephalopathy (SAE) manifests clinically in hyperneuroinflammation. Pyroptosis, which can induce an inflammatory cascade response, has been considered to be a causative factor of SAE. Evidence has shown that the bromo- and extraterminal (BET) proteins (including BRD2, BRD3, BRD4 and BRDT) inhibitor JQ1 can inhibit inflammation and suppress pyroptosis in various diseases. Therefore, we examined the effect of JQ1 on inflammasome-induced pyroptosis in the hippocampus in a mouse model of sepsis induced by lipopolysaccharide (LPS) injection. The results showed that JQ1 treatment alleviated sepsis-related symptoms, protected the blood–brain barrier (BBB), as indicated by upregulated expression of the tight junction proteins occludin and ZO-1, and remarkably rescued neuronal damage in SAE mice. Mechanistically, we demonstrated that JQ1 intervention inhibited the expression of BRD proteins and decreased the expression of inflammasomes by blocking nuclear factor kappa B (NFκB) signalling, attenuating the canonical pyroptosis (mediated by cleaved-Caspase1/11) pathway and the release of proinflammatory factors in the hippocampus of septic mice. Interestingly, we also found that JQ1 selectively suppressed the activation of hippocampal microglia in SAE mice. Thus, JQ1 protected the hippocampal BBB and neuronal damage through the attenuation of neuroinflammation by inhibiting the inflammasome-dependent canonical pyroptosis pathway induced by LPS injection in mice, and JQ1 may be a promising target for the prevention of SAE.
- Published
- 2022
37. Expression of nucleolin, nucleophosmin, upstream binding transcription factor genes and propolis in wound models
- Author
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Murat, Kaya, Recep, Eroz, Murat, Kabakliogli, and Tıp Fakültesi
- Subjects
Nucleolar Proteins ,Nucleolin ,Antifungal Agents ,Nursing (miscellaneous) ,Nuclear Proteins ,RNA-Binding Proteins ,Gene Expression ,Phosphoproteins ,Antifungal ,Antiviral Agents ,Propolis ,Anti-Bacterial Agents ,Upstream Binding Transcription Factors ,Antibacterial ,Animals ,Fundamentals and skills ,Antiviral ,Bee Propolis ,Nucleophosmin ,Transcription Factors - Abstract
Objective: Nucleolar proteins have important functions in the regulation of cell homeostasis and play a crucial role in sensing various types of stress, such as genotoxic stress. Propolis has epithelial, analgesic, antibacterial, antifungal and antiviral effects. This study aimed to evaluate the gene expression levels of nucleolar proteins: nucleolin (NCL); nucleophosmin (NPM1); and upstream binding transcription factor (UBTF), as well as the benefits of propolis in wound healing. Method: This experimental study was conducted by creating clean and clean–contaminated wounds according to the Surgical Site Infection Guidelines, 2016. A total of seven animal groups were included in the study: control; laparotomy; anastomosis; fucidic acid with/without anastomosis; propolis with/without anastomosis Results: Statistically significant differences of levels of gene expression among the groups were detected for NCL (p=0.004), NPM1 (p=0.011) and UBTF (p=0.000). When the expression levels of the related genes and blood parameters are considered, the relationship between NCL, NPM1 and UBTF expression levels and blood parameters (NE, EO, NE(%), LY, LY(%), EO, EO(%), MO, MO(%), RBC, HB, HCT, MCV, MCHC, RDW, RDW(%), PLT, PDW and PCT) were statistically significant. Conclusion: The nucleolar proteins such as NCL, NPM1 and UBTF have important functions in cell viability and its maintenance under various condition such as stress and injury. Additionally, propolis has positive benefits in wound healing and in the prevention of wound infection. Our findings provide the first insights into the putative role of those proteins in wound healing.
- Published
- 2022
38. An In Vivo Model for Elucidating the Role of an Erythroid-Specific Isoform of Nuclear Export Protein Exportin 7 (Xpo7) in Murine Erythropoiesis
- Author
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Susree Modepalli, Sandra Martinez-Morilla, Srividhya Venkatesan, James Fasano, Katerina Paulsen, Dirk Görlich, Shilpa Hattangadi, and Gary M. Kupfer
- Subjects
Mice ,Cancer Research ,ran GTP-Binding Protein ,Active Transport, Cell Nucleus ,Genetics ,Animals ,Nuclear Proteins ,Protein Isoforms ,Erythropoiesis ,Cell Biology ,Hematology ,Karyopherins ,Molecular Biology - Abstract
Erythroid nuclear condensation is a complex process in which compaction to one-tenth its original size occurs in an active nucleus simultaneously undergoing transcription and cell division. We previously found that the nuclear exportin Exportin7 (Xpo7), which is erythroid- specific and highly induced during terminal erythropoiesis, facilitates nuclear condensation. We also identified a previously unannotated, erythroid-specific isoform of Xpo7 (Xpo7B) containing a novel first exon Xpo7-1b expressed only in late Ter119
- Published
- 2022
39. Excessive activation of HOXB13/PIMREG axis promotes hepatocellular carcinoma progression and drug resistance
- Author
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Cui, Tang, Shixiong, Qiu, Wenying, Mou, Jinming, Xu, and Peijun, Wang
- Subjects
Homeodomain Proteins ,Carcinoma, Hepatocellular ,Liver Neoplasms ,Drug Resistance ,Intracellular Signaling Peptides and Proteins ,Biophysics ,Down-Regulation ,Nuclear Proteins ,Cell Biology ,Protein Serine-Threonine Kinases ,Biochemistry ,Gene Expression Regulation, Neoplastic ,Cell Line, Tumor ,Humans ,Carrier Proteins ,Molecular Biology ,Cell Proliferation ,Transcription Factors - Abstract
The transcription factor HOXB13 is bound up with the occurrence, progression and drug fast of many kinds of cancer. Nevertheless, the specific molecular mechanism of HOXB13 in hepatocellular carcinoma (HCC) is still unknown. This provides an obstacle to the exploration of HCC treatments targeting HOXB13. This study found that HOXB13 was up-regulated in HCC tissues. HOXB13 enhanced the multiplication and metastasis of HCC cells. It enhanced HCC cell drug and anoikis resistance. The analysis of HCC RNA seq data indicated that the expression of HOXB13 and PIMREG were positively correlated. Luciferase report assay showed that HOXB13 could activate PIMREG promoter transcription. The results of RT-qPCR and western blot showed that HOXB13 regulated the transcription of PIMREG. Western blot proved that high expression of PIMREG participated in DNA damage repair and cell cycle regulation by up-regulating RAD51, BRCA1, CDC25A, CDC25B and CDC25C and down-regulating HIPK2. This led to a significant increase in DNA repair capacity, accelerated cell cycle progression, and insensitive to DNA damage. Down-regulation of PIMREG in Hep3B cells overexpressing HOXB13 attenuated the phenotype induced by HOXB13. Therefore, HOXB13 functioned through PIMREG instead of directly regulating the transcription of RAD51, BRCA1, CDC25A, CDC25B and CDC25C. The same results were obtained in vivo. It was concluded that HOXB13 affected the expression of cell cycle and DNA repair related factors by up-regulating the transcription of PIMREG, thereby promoting the progression of HCC and enhancing the resistance of HCC to chemotherapeutics.
- Published
- 2022
40. Analysis of Slow-Cycling Variants of the Light-Inducible Nuclear Protein Export System LEXY in Mammalian Cells
- Author
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Forlani, Giada, Antwi, Enoch B., Weis, Daniel, Öztürk, Mehmet A., Queck, Bastian A.W., Brecht, Dominik, and Di Ventura, Barbara
- Subjects
Nuclear Export Signals ,Oxygen ,Mammals ,Phototropins ,Tumor Necrosis Factor Ligand Superfamily Member 14 ,Light ,Avena ,Biomedical Engineering ,Animals ,Nuclear Proteins ,General Medicine ,Biochemistry, Genetics and Molecular Biology (miscellaneous) - Abstract
The optogenetic tool LEXY consists of the second light oxygen voltage (LOV) domain of
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- 2022
41. KDOAM-25 Overcomes Resistance to MEK Inhibitors by Targeting KDM5B in Uveal Melanoma
- Author
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Hongjun Zhang, Xiangnan Liu, Yong’an Chen, Rui Xu, and Shengli He
- Subjects
Mitogen-Activated Protein Kinase Kinases ,Niacinamide ,Uveal Neoplasms ,Jumonji Domain-Containing Histone Demethylases ,Article Subject ,General Immunology and Microbiology ,Annexins ,Lysine ,Glycine ,Nuclear Proteins ,General Medicine ,General Biochemistry, Genetics and Molecular Biology ,Epigenesis, Genetic ,Histones ,Repressor Proteins ,Cell Line, Tumor ,Humans ,Melanoma ,Protein Kinase Inhibitors ,Proto-Oncogene Proteins c-akt ,Cell Proliferation - Abstract
Background. Uveal Melanoma (UM) is a potentially lethal cancer, and epigenetics may participate in the regulation of MEK resistance. This study is aimed at targeting the epigenetic kinase to overcome the resistance to MEK inhibitor. Method. We developed the 92.1 and OMM1 MEK-inhibitor resistant cell lines by culturing them in the trametinib (Tra) mixed medium. We utilized CCK8 analysis for detecting the viability of the cell. Western blot was used to determine the ERK1/2 and Akt phosphorylation. Small compound library screening assays were carried out by CCK8 analysis. To test the apoptosis, we employed flow cytometric analysis with Annexin-V/PI. Western blot and CCK8 were used to explore the epigenetic regulation of KDM5B in MEK-resistance cell lines. To knock out the expression level of KDM5B, we used the CRISPR/Cas9 by lentivirus delivering well-validated shRNAs in pLKO.1 vector. The directly binding affinity of KDOAM-25 to KDM5B was determined by drug affinity responsive target stability (DARTS) and microscale thermophoresis (MST). Results. The phosphorylation of ERK1/2 and Akt (T308) was inhibited in OMM1 cell lines. However, inhibition of Tra was abolished in OMM1-R cell lines. From a compound screening assay, we identified that KDOAM-25 robustly inhibited the viability and colony formation of MEK-resistance cell lines. Furthermore, KDOAM-25 significantly promoted cell death in OMM1-R cells. H3K4me3 (tri-methylation of lysine 4 on histone H3) and H3K27ac (acetyl of lysine 27 on histone H3) were both upregulated in OMM1-R cells. Tra significantly inhibited the expression of KDM5B in OMM1-P cells. However, the effect on KDM5B was abolished in OMM1-R cells. Knockdown of KDM5B robustly suppressed the cell viability in OMM1-R cells. KDOAM-25 directly interacted with KDM5B. Conclusion. KDOAM-25 inhibited the viability and colony formation and promoted cell death of MEK-resistance cell lines through H3K4me3 and H3K27ac, indicating that KDOAM-25 may be a potential therapeutic agent for MEK resistance in UM patients.
- Published
- 2022
42. Diagnosis, Treatment and Prognosis of Primary Pulmonary NUT Carcinoma: A Literature Review
- Author
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Jiaqian Yuan, Zhili Xu, and Yong Guo
- Subjects
Adult ,Male ,Lung Neoplasms ,Carcinoma ,Humans ,Nuclear Proteins ,Prognosis ,Neoplasm Proteins - Abstract
NUT carcinoma is a rare, highly lethal cancer characterized with the rearrangement of the nuclear protein in testis (NUT) gene on chromosome 15q14, which primarily occurs in the midline organs. Primary pulmonary NUT carcinoma (NC) lacks characteristic clinical manifestations, which leads to the high rate of misdiagnose and nonstandard treatment. To date, fewer than one hundred cases have been reported worldwide. Here, a comprehensive literature search involving a total of 35 articles with 55 patients was conducted in this paper. We reviewed and analyzed the associated clinical and pathological characteristics, the efficacy of various treatment options and the prognosis. Pulmonary NC mainly occurred in middle young-aged men (median age, 36) with no smoking history (2:1) and would present with symptoms of cough (63.6%), dyspnea (29.5%), chest pain (18.2%) and hemoptysis (18.2%). The initial imaging frequently revealed large and irregular lesions in the lower lobe (46.5%) of the left or right lungs; lymph node metastasis was also prevalent (91.9%). A focal squamous differentiation with abrupt keratinization often occurred in the undifferentiated or poorly differentiated (93.75%) tumor cells, with abundant necrosis and numerous neutrophils infiltrated. The mean overall survival (OS) in patients of this malignant disease was 6.21 months, and the median OS was 4.4 months. According to our results, this disease is sensitive to radiotherapy, and chemoradiotherapy (either concurrent chemoradiotherapy or sequential chemoradiotherapy) was the most efficient therapeutic regimen to prolong the OS of patients with pulmonary NC.
- Published
- 2022
43. FIGNL1 Expression and its Prognostic Significance in Pan-cancer Analysis
- Author
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Minghao Li, Zicheng Zhen, Muyan Zhong, Liqun Ye, and Xiaofang Ma
- Subjects
Adenosine Triphosphatases ,Carcinoma, Hepatocellular ,Liver Neoplasms ,Organic Chemistry ,Nuclear Proteins ,AAA Proteins ,General Medicine ,Prognosis ,Computer Science Applications ,Drug Discovery ,ATPases Associated with Diverse Cellular Activities ,Humans ,Microtubule-Associated Proteins ,Biomarkers - Abstract
Background: Fidgetin-like 1 (FIGNL1), a subfamily member of ATPases, is associated with diverse cellular activities (AAA proteins). FIGNL1 is involved in DNA repair. However, the latest study has indicated that FIGNL1 plays a crucial role in the occurrence and development of malignant tumors. Methods: FIGNL1 expression was analyzed via Oncomine and GEPIA databases, and its prognostic potential was analyzed using OncoLnc, UALCAN, and GEPIA databases. Moreover, the promoter methylation of FIGNL1 was analyzed through the UALCAN database. FIGNL1-related gene network was found within STRING. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were investigated across WebGestalt. FIGNL1 correlation with cancer immune infiltrates was estimated using the Tumor Immune Estimation Resource (TIMER) database. Results: We found that FIGNL1 is widely overexpressed in multiple human cancers, and its high expression was correlated with the poor prognosis of patients with kidney renal clear-cell carcinoma (KIRP), low-grade glioma (LGG) of brain and liver hepatocellular carcinoma (LIHC). Additionally, the promoter methylation level of FIGNL1 showed a statistical significance between normal and primary tissues in KIRP and LGG via the UALCAN (P < 0.0001). FIGNL1 expression was highly correlated with the infiltrating levels of CD8+ T and CD4+ T cells, dendritic cells (DCs), macrophages, and neutrophils in LIHC. Conclusions: In this study, the correlation of FIGNL1 expression with the prognosis, promoter methylation, and immune infiltrates in KIRP, LGG, and LIHC was revealed. These findings suggested that FIGNL1 promised to be a prognostic biomarker for KIRP, LGG, and LIHC.
- Published
- 2022
44. Preliminary Evaluation of Artificial Intelligence-Based Anti-Hepatocellular Carcinoma Molecular Target Study in Hepatocellular Carcinoma Diagnosis Research
- Author
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Yuan Wang, Chao Wei, Xiangui Deng, Shudi Gao, and Jing Chen
- Subjects
Carcinoma, Hepatocellular ,General Immunology and Microbiology ,Artificial Intelligence ,Liver Neoplasms ,Humans ,Nuclear Proteins ,Cell Cycle Proteins ,General Medicine ,General Biochemistry, Genetics and Molecular Biology ,Transcription Factors - Abstract
In this paper, in-depth research analysis of anti-hepatocellular carcinoma molecular targets for hepatocellular carcinoma diagnosis was conducted using artificial intelligence. Because BRD4 plays an important role in gene transcription for cell cycle regulation and apoptosis, tumor-targeted therapy by inhibiting the expression or function of BRD4 has received increasing attention in the field of antitumor research. Study subjects in small samples were used as the validation set for validating each diagnostic model constructed based on the training set. The diagnostic effect of each model in the validation set is evaluated by calculating the sensitivity, specificity, and compliance rate, and the model with the best and most stable diagnostic value is selected by combining the results of model construction, validation, and evaluation. The total sample was divided into a training set and test set by using a stratified sampling method in the ratio of 7 : 3. Logistic regression, weighted k -nearest neighbor, decision tree, and BP artificial neural network were used in the training set to construct diagnostic models for early-stage liver cancer, respectively, and the optimal parameters of the corresponding models were obtained, and then, the constructed models were validated in the test set. To evaluate the diagnostic efficacy, stability, and generalization ability of the four classification methods more robustly, a 10-fold crossover test was performed for each classification method. BRD4 is an epigenetic regulator that is associated with the upregulation of expression of various oncogenic drivers in tumors. Targeting BRD4 with pharmacological inhibitors has emerged as a novel approach for tumor treatment. However, before we implemented this topic, there were no detailed studies on whether BRD4 could be used for the treatment of HCC, the role of BRD4 in HCC cell proliferation and apoptosis, and the ability of small molecule BRD4 inhibitors to induce apoptosis in hepatocellular carcinoma cells.
- Published
- 2022
45. The high mobility group protein HMG20A cooperates with the histone reader PHF14 to modulate TGFβ and Hippo pathways
- Author
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Elena Gómez-Marín, Melanija Posavec-Marjanović, Laura Zarzuela, Laura Basurto-Cayuela, José A Guerrero-Martínez, Gonzalo Arribas, Rosario Yerbes, María Ceballos-Chávez, Manuel Rodríguez-Paredes, Mercedes Tomé, Raúl V Durán, Marcus Buschbeck, José C Reyes, Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), European Commission, Junta de Andalucía, Fundación Vencer el Cancer, Fundació La Marató de TV3, and Generalitat de Catalunya
- Subjects
Proteomics ,High Mobility Group Proteins ,Nuclear Proteins ,Cadherins ,Chromatin ,Histones ,Transforming Growth Factor beta ,Cell Line, Tumor ,Genetics ,Humans ,Hippo Signaling Pathway ,RNA, Small Interfering ,Transcription Factors - Abstract
High mobility group (HMG) proteins are chromatin regulators with essential functions in development, cell differentiation and cell proliferation. The protein HMG20A is predicted by the AlphaFold2 software to contain three distinct structural elements, which we have functionally characterized: i) an amino-terminal, intrinsically disordered domain with transactivation activity; ii) an HMG box with higher binding affinity for double-stranded, four-way-junction DNA than for linear DNA; and iii) a long coiled-coil domain. Our proteomic study followed by a deletion analysis and structural modeling demonstrates that HMG20A forms a complex with the histone reader PHF14, via the establishment of a two-stranded alpha-helical coiled-coil structure. siRNA-mediated knockdown of either PHF14 or HMG20A in MDA-MB-231 cells causes similar defects in cell migration, invasion and homotypic cell–cell adhesion ability, but neither affects proliferation. Transcriptomic analyses demonstrate that PHF14 and HMG20A share a large subset of targets. We show that the PHF14-HMG20A complex modulates the Hippo pathway through a direct interaction with the TEAD1 transcription factor. PHF14 or HMG20A deficiency increases epithelial markers, including E-cadherin and the epithelial master regulator TP63 and impaired normal TGFβ-trigged epithelial-to-mesenchymal transition. Taken together, these data indicate that PHF14 and HMG20A cooperate in regulating several pathways involved in epithelial–mesenchymal plasticity., Research in the J.C. Reyes lab was funded by the Spanish Ministry of Economy and Competitiveness MCIN/AEI/10.13039/501100011033/ [PID2020-118516GB-I00]; Junta de Andalucía [P18-FR-1962 and BIO-321]; Fundación Vencer El Cancer (VEC); European Union FEDER ‘A way to build Europe’ program; research at Buschbeck lab is further supported by FEDER/Ministerio de Ciencia e Innovación - Agencia Estatal de Investigación [RTI2018-094005-B-I00]; Marie Skłodowska Curie Training network ‘INTERCEPT-MDS’ [H2020-MSCA-ITN-2020–953407]; Deutsche José Carreras Leukämie Stiftung [DJCLS 14R/2018, AGAUR 2017-SGR-305]; Fundació La Marató de TV3 257/C/2019; CABIMER is a Center partially funded by the Junta de Andalucía; E.G.-M. is the recipient of an FPI fellowship from the Spanish Ministry of Science and Innovation; L.B.-C. is the recipient of an FPU fellowship from the Spanish Ministry of Education; M.P.M. was funded by an AGAUR FI-2010 fellowship. Funding for open access charge: Spanish Ministry of Economy and Competitiveness MCIN/AEI/10.13039/501100011033/ [PID2020-118516GB-I00].
- Published
- 2022
46. Identification of Histone Peptide Binding Specificity and Small-Molecule Ligands for the TRIM33α and TRIM33β Bromodomains
- Author
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Angelina R. Sekirnik, Jessica K. Reynolds, Larissa See, Joseph P. Bluck, Amy R. Scorah, Cynthia Tallant, Bernadette Lee, Katarzyna B. Leszczynska, Rachel L. Grimley, R. Ian Storer, Marta Malattia, Sara Crespillo, Sofia Caria, Stephanie Duclos, Ester M. Hammond, Stefan Knapp, Garrett M. Morris, Fernanda Duarte, Philip C. Biggin, and Stuart J. Conway
- Subjects
Histones ,DNA-Binding Proteins ,Peptide T ,Lysine ,Ubiquitin-Protein Ligases ,Nuclear Proteins ,Molecular Medicine ,General Medicine ,Ligands ,Ubiquitins ,Biochemistry ,Transcription Factors - Abstract
TRIM33 is a member of the tripartite motif (TRIM) family of proteins, some of which possess E3 ligase activity and are involved in the ubiquitin-dependent degradation of proteins. Four of the TRIM family proteins, TRIM24 (TIF1α), TRIM28 (TIF1β), TRIM33 (TIF1γ) and TRIM66, contain C-terminal plant homeodomain (PHD) and bromodomain (BRD) modules, which bind to methylated lysine (KMen) and acetylated lysine (KAc), respectively. Here we investigate the differences between the two isoforms of TRIM33, TRIM33α and TRIM33β, using structural and biophysical approaches. We show that the N1039 residue, which is equivalent to N140 in BRD4(1) and which is conserved in most BRDs, has a different orientation in each isoform. In TRIM33β, this residue coordinates KAc, but this is not the case in TRIM33α. Despite these differences, both isoforms show similar affinities for H31–27K18Ac, and bind preferentially to H31–27K9Me3K18Ac. We used this information to develop an AlphaScreen assay, with which we have identified four new ligands for the TRIM33 PHD-BRD cassette. These findings provide fundamental new information regarding which histone marks are recognized by both isoforms of TRIM33 and suggest starting points for the development of chemical probes to investigate the cellular function of TRIM33.
- Published
- 2022
47. Splicing of branchpoint-distant exons is promoted by Cactin, Tls1 and the ubiquitin-fold-activated Sde2
- Author
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Anupa T Anil, Karan Choudhary, Rakesh Pandian, Praver Gupta, Poonam Thakran, Arashdeep Singh, Monika Sharma, and Shravan Kumar Mishra
- Subjects
Ubiquitin ,RNA Splicing ,Telomere-Binding Proteins ,Nuclear Proteins ,Exons ,Ribonucleoprotein, U2 Small Nuclear ,Introns ,Shelterin Complex ,DNA-Binding Proteins ,Alternative Splicing ,Heterochromatin ,Schizosaccharomyces ,RNA Precursors ,Genetics ,Humans ,RNA Splice Sites ,Schizosaccharomyces pombe Proteins ,Carrier Proteins - Abstract
Intron diversity facilitates regulated gene expression and alternative splicing. Spliceosomes excise introns after recognizing their splicing signals: the 5′-splice site (5′ss), branchpoint (BP) and 3′-splice site (3′ss). The latter two signals are recognized by U2 small nuclear ribonucleoprotein (snRNP) and its accessory factors (U2AFs), but longer spacings between them result in weaker splicing. Here, we show that excision of introns with a BP-distant 3′ss (e.g. rap1 intron 2) requires the ubiquitin-fold-activated splicing regulator Sde2 in Schizosaccharomyces pombe. By monitoring splicing-specific ura4 reporters in a collection of S. pombe mutants, Cay1 and Tls1 were identified as additional regulators of this process. The role of Sde2, Cay1 and Tls1 was further confirmed by increasing BP–3′ss spacings in a canonical tho5 intron. We also examined BP-distant exons spliced independently of these factors and observed that RNA secondary structures possibly bridged the gap between the two signals. These proteins may guide the 3′ss towards the spliceosome's catalytic centre by folding the RNA between the BP and 3′ss. Orthologues of Sde2, Cay1 and Tls1, although missing in the intron-poor Saccharomyces cerevisiae, are present in intron-rich eukaryotes, including humans. This type of intron-specific pre-mRNA splicing appears to have evolved for regulated gene expression and alternative splicing of key heterochromatin factors.
- Published
- 2022
48. DSC2 Suppresses the Metastasis of Gastric Cancer through Inhibiting the BRD4/Snail Signaling Pathway and the Transcriptional Activity of β-Catenin
- Author
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Chao Sun, Lei Wang, Dan-dan Du, Jian-bo Ji, Xiao-xia Yang, Bing-fang Yu, Peng-fei Shang, and Xiu-Li Guo
- Subjects
Desmocollins ,Aging ,Article Subject ,Nuclear Proteins ,Cell Cycle Proteins ,Cell Biology ,General Medicine ,Biochemistry ,Gene Expression Regulation, Neoplastic ,Mice ,Stomach Neoplasms ,Cell Line, Tumor ,Animals ,Humans ,beta Catenin ,Signal Transduction ,Transcription Factors - Abstract
Downregulated DSC2 involved in the metastasis of cancers. Unfortunately, its role on the development of gastric cancer (GC) and the potential mechanisms remain unclear. Bioinformatics analysis, Western blot, qRT-PCR, and immunohistochemistry were performed to detect the DSC2 levels of human GC and normal stomach tissues. The role of DSC2 and the downstream signaling in gastric carcinogenesis were explored by using GC specimens, GC cells with different DSC2 expression, inhibitors, and mouse metastasis models. We found that the level of DSC2 decreased significantly in GC tissues and cells. Recovered DSC2 inhibited the invasion and migration of GC cells both in culture and in xenografts. Mechanistically, DSC2 could not only decrease Snail level and nuclear BRD4 level by forming DSC2/BRD4, but also inhibit nuclear translocation of β-catenin. We concluded that DSC2 inhibited the metastasis of GC, and the underlying mechanisms were closely related to the regulation on nuclear translocation of BRD4 and β-catenin. Our results suggest that DSC2 may serve as a novel therapeutic target for GC.
- Published
- 2022
49. Nested epistasis enhancer networks for robust genome regulation
- Author
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Xueqiu Lin, Yanxia Liu, Shuai Liu, Xiang Zhu, Lingling Wu, Yanyu Zhu, Dehua Zhao, Xiaoshu Xu, Augustine Chemparathy, Haifeng Wang, Yaqiang Cao, Muneaki Nakamura, Jasprina N. Noordermeer, Marie La Russa, Wing Hung Wong, Keji Zhao, and Lei S. Qi
- Subjects
Machine Learning ,Enhancer Elements, Genetic ,Multidisciplinary ,Humans ,Nuclear Proteins ,Cell Cycle Proteins ,Disease ,Epistasis, Genetic ,K562 Cells ,Article ,Genome-Wide Association Study ,Transcription Factors - Abstract
Mammalian genomes possess multiple enhancers spanning an ultralong distance (>megabases) to modulate important genes, yet it is unclear how these enhancers coordinate to achieve this task. Here, we combine multiplexed CRISPRi screening with machine learning to define quantitative enhancer-enhancer interactions. We find that the ultralong distance enhancer network possesses a nested multi-layer architecture that confers functional robustness of gene expression. Experimental characterization reveals that enhancer epistasis is maintained by three-dimensional chromosomal interactions and BRD4 condensation. Machine learning prediction of synergistic enhancers provides an effective strategy to identify non-coding variant pairs associated with pathogenic genes in diseases beyond Genome-Wide Association Studies (GWAS) analysis. Our work unveils nested epistasis enhancer networks, which can better explain enhancer functions within cells and in diseases.
- Published
- 2022
50. Unified gene expression signature of novel NPM1 exon 5 mutations in acute myeloid leukemia
- Author
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Véronique Lisi, Ève Blanchard, Michael Vladovsky, Éric Audemard, Albert Ferghaly, Sébastien Lemieux, Josée Hébert, Guy Sauvageau, and Vincent-Philippe Lavallée
- Subjects
Leukemia, Myeloid, Acute ,Mutation ,Humans ,Nuclear Proteins ,Exons ,Hematology ,Transcriptome - Published
- 2022
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