79 results on '"Presutti C"'
Search Results
2. microRNAs Modulate Spatial Memory in the Hippocampus and in the Ventral Striatum in a Region-Specific Manner
- Author
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Capitano, F., Camon, J., Ferretti, V., Licursi, V., De Vito, F., Rinaldi, A., Vincenti, S., Mannironi, C., Fragapane, P., Bozzoni, I., Oliverio, A., Negri, R., Presutti, C., and Mele, Andrea
- Published
- 2016
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3. MicroRNA-335-5p modulates spatial memory and hippocampal synaptic plasticity
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Capitano, F., Camon, J., Licursi, V., Ferretti, V., Maggi, L., Scianni, M., Del Vecchio, G., Rinaldi, A., Mannironi, C., Limatola, C., Presutti, C., and Mele, A.
- Published
- 2017
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4. Epstein–Barr virus encoded LMP1 downregulates TCL1 oncogene through miR-29b
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Anastasiadou, E, Boccellato, F, Vincenti, S, Rosato, P, Bozzoni, I, Frati, L, Faggioni, A, Presutti, C, and Trivedi, P
- Published
- 2010
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5. Separation of Structurally Related Anthocyanins by MEKC
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Bednar, P., Tomassi, A.V., Presutti, C., Pavlikova, M., Lemr, K., and Fanali, S.
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- 2003
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6. Differential regulation of miR-21 and miR-146a by Epstein-Barr virus-encoded EBNA2
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Rosato, P, Anastasiadou, E, Garg, N, Lenze, D, Boccellato, F, Vincenti, S, Severa, M, Coccia, E M, Bigi, R, Cirone, M, Ferretti, E, Campese, A F, Hummel, M, Frati, L, Presutti, C, Faggioni, A, and Trivedi, P
- Published
- 2012
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7. Identification of a regulated pathway for nuclear pre-mRNA turnover
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Bousquet-Antonelli, C., Presutti, C., and Tollervey, D.
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Cytochemistry -- Research ,Messenger RNA -- Physiological aspects ,Cellular signal transduction -- Genetic aspects ,Cell nuclei -- Physiological aspects ,Yeast -- Genetic aspects ,Gene expression -- Research ,Genetic regulation -- Physiological aspects ,Enzymes -- Physiological aspects ,Biological sciences - Abstract
A nuclear pathway that quickly degrades unspliced pre-mRNAs in yeast has been identified. It involves 3'-to-5' degradation by the exosome complex and 5'-to-3' degradation by the exonuclease Rat1p. It is proposed that nuclear pre-mRNA turnover is a novel step in gene expression regulation.
- Published
- 2000
8. One-step nested PCR for detection of 2 LTR circles in PBMCs of HIV-1 infected patients with no detectable plasma HIV RNA
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Lusi, E.A., Guarascio, P., Presutti, C., Villani, R., Pellicelli, A., and Soccorsi, F.
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- 2005
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9. Ecotoxicological effects of a nonionic detergent (Triton DF-16) assayed by ModFETAX
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Presutti, C., Vismara, C., Camatini, M., and Bernardini, G.
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- 1994
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10. Evaluation of the developmental toxicity of the pesticide MCPA and its contaminants phenol and chlorocresol
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Bernardini, G., Vismara, C., Orlandi, M., Bolzacchini, E., Presutti, C., Spinelli, O., and Settimi, R.
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CHLOROCRESOL ,PHENOL ,AMPHIBIANS ,PESTICIDES - Published
- 1996
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11. The reactivity and fate of a phenoxyacetic herbicide: 4-chloro-2-methylphenoxyacetic acid (MCPA) in the environment
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Bolzacchini, E., Orlandi, M., Presutti, C., Bernardini, G., and Rindone, B.
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HERBICIDES - Published
- 1995
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12. Epstein-Barr virus infection leads to partial phenotypic reversion of terminally differentiated malignant B cells.
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Anastasiadou E, Vaeth S, Cuomo L, Boccellato F, Vincenti S, Cirone M, Presutti C, Junker S, Winberg G, Frati L, Wade PA, Faggioni A, Trivedi P, Anastasiadou, Eleni, Vaeth, Signe, Cuomo, Laura, Boccellato, Francesco, Vincenti, Sara, Cirone, Mara, and Presutti, Carlo
- Abstract
The B cell lymphomas associated with Epstein-Barr virus (EBV) are not limited to any specific stage of B cell differentiation but covers widely different B cell phenotypes. In vitro infection of the virus negative tumors with a recombinant EBV strain has provided important insights into virus-tumor interaction. Here, we investigated the interaction between EBV and terminally differentiated tumor derived B cells, namely multiple myeloma (MM). The in vitro EBV infected MM expressed restricted viral latency. Acquisition of the virus was accompanied by a partial reprogramming to a mature B cell phenotype. Thus, the plasma cell markers syndecan-1 (CD138), Blimp1 and MUM1 were downregulated, while expression of HLADR, CIITA and TCL1, which are normally not expressed in plasmacytoid cells, was upregulated. The silenced transcription factor gene encoding Pax5 and its target BLNK were activated. Significantly, the free lambda light chains secreted in the medium were reduced in EBV infected MM clones. Collectively, these results suggest that the restricted EBV latency can cause at least partial phenotypic reversion of terminally differentiated B tumor cells. We suggest that the restricted EBV latent gene expression may not only be the consequence but the cause of the mature B cell phenotype, actively participating in the virus persistence. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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13. Reversing vemurafenib-resistance in primary melanoma cells by combined romidepsin and type I IFN treatment through blocking of tumorigenic signals and induction of immunogenic effects.
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Fragale A, Stellacci E, Romagnoli G, Licursi V, Parlato S, Canini I, Remedi G, Buoncervello M, Matarrese P, Pedace L, Ascione B, Pizzi S, Tartaglia M, D'Atri S, Presutti C, Capone I, and Gabriele L
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- Humans, Vemurafenib pharmacology, Vemurafenib therapeutic use, Drug Resistance, Neoplasm, Protein Kinase Inhibitors pharmacology, Proto-Oncogene Proteins B-raf genetics, Cell Line, Tumor, Melanoma drug therapy, Melanoma genetics, Melanoma pathology, Interferon Type I
- Abstract
BRAF
V600 mutations are the most common oncogenic alterations in melanoma cells, supporting proliferation, invasion, metastasis and immune evasion. In patients, these aberrantly activated cellular pathways are inhibited by BRAFi whose potent antitumor effect and therapeutic potential are dampened by the development of resistance. Here, by using primary melanoma cell lines, generated from lymph node lesions of metastatic patients, we show that the combination of two FDA-approved drugs, the histone deacetylate inhibitor (HDCAi) romidepsin and the immunomodulatory agent IFN-α2b, reduces melanoma proliferation, long-term survival and invasiveness and overcomes acquired resistance to the BRAFi vemurafenib (VEM). Targeted resequencing revealed that each VEM-resistant melanoma cell line and the parental counterpart are characterized by a distinctive and similar genetic fingerprint, shaping the differential and specific antitumor modulation of MAPK/AKT pathways by combined drug treatment. By using RNA-sequencing and functional in vitro assays, we further report that romidepsin-IFN-α2b treatment restores epigenetically silenced immune signals, modulates MITF and AXL expression and induces both apoptosis and necroptosis in sensitive and VEM-resistant primary melanoma cells. Moreover, the immunogenic potential of drug-treated VEM-resistant melanoma cells results significantly enhanced, given the increased phagocytosis rate of these cells by dendritic cells, which in turn exhibit also a selective down-modulation of the immune checkpoint TIM-3. Overall, our results provide evidence that combined epigenetic-immune drugs can overcome VEM resistance of primary melanoma cells by oncogenic and immune pathways reprogramming, and pave the way for rapidly exploiting this combination to improve BRAFi-resistant metastatic melanoma treatment, also via reinforcement of immune checkpoint inhibitor therapy., (© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.)- Published
- 2023
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14. Rational design and synthesis of a novel BODIPY-based probe for selective imaging of tau tangles in human iPSC-derived cortical neurons.
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Soloperto A, Quaglio D, Baiocco P, Romeo I, Mori M, Ardini M, Presutti C, Sannino I, Ghirga S, Iazzetti A, Ippoliti R, Ruocco G, Botta B, Ghirga F, Di Angelantonio S, and Boffi A
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- Boron Compounds, Humans, Neurons metabolism, tau Proteins metabolism, Induced Pluripotent Stem Cells metabolism
- Abstract
Numerous studies have shown a strong correlation between the number of neurofibrillary tangles of the tau protein and Alzheimer's disease progression, making the quantitative detection of tau very promising from a clinical point of view. However, the lack of highly reliable fluorescent probes for selective imaging of tau neurofibrillary tangles is a major challenge due to sharing similar β-sheet motifs with homologous Amyloid-β fibrils. In the current work, we describe the rational design and the in silico evaluation of a small-size focused library of fluorescent probes, consisting of a BODIPY core (electron acceptor) featuring highly conjugated systems (electron donor) with a length in the range 13-19 Å at C3. Among the most promising probes in terms of binding mode, theoretical affinity and polarity, BT1 has been synthesized and tested in vitro onto human induced pluripotent stem cells derived neuronal cell cultures. The probe showed excellent photophysical properties and high selectivity allowing in vitro imaging of hyperphosphorylated tau protein filaments with minimal background noise. Our findings offer new insight into the structure-activity relationship of this class of tau selective fluorophores, paving the way for boosting tau tangle detection in patients possibly through retinal spectral scans., (© 2022. The Author(s).)
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- 2022
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15. The microRNA let-7b-5p Is Negatively Associated with Inflammation and Disease Severity in Multiple Sclerosis.
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Mandolesi G, Rizzo FR, Balletta S, Stampanoni Bassi M, Gilio L, Guadalupi L, Nencini M, Moscatelli A, Ryan CP, Licursi V, Dolcetti E, Musella A, Gentile A, Fresegna D, Bullitta S, Caioli S, Vanni V, Sanna K, Bruno A, Buttari F, Castelli C, Presutti C, De Santa F, Finardi A, Furlan R, Centonze D, and De Vito F
- Subjects
- Adult, Female, Humans, Male, Middle Aged, Multiple Sclerosis pathology, Inflammation metabolism, MicroRNAs metabolism, Multiple Sclerosis genetics
- Abstract
The identification of microRNAs in biological fluids for diagnosis and prognosis is receiving great attention in the field of multiple sclerosis (MS) research but it is still in its infancy. In the present study, we observed in a large sample of MS patients that let-7b-5p levels in the cerebrospinal fluid (CSF) were highly correlated with a number of microRNAs implicated in MS, as well as with a variety of inflammation-related protein factors, showing specific expression patterns coherent with let-7b-5p-mediated regulation. Additionally, we found that the CSF let-7b-5p levels were significantly reduced in patients with the progressive MS compared to patients with relapsing-remitting MS and were negatively correlated with characteristic hallmark processes of the two phases of the disease. Indeed, in the non-progressive phase, let-7b-5p inversely associated with both central and peripheral inflammation; whereas, in progressive MS, the CSF levels of let-7b-5p negatively correlated with clinical disability at disease onset and after a follow-up period. Overall, our results uncovered, by the means of a multidisciplinary approach and multiple statistical analyses, a new possible pleiotropic action of let-7b-5p in MS, with potential utility as a biomarker of MS course.
- Published
- 2021
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16. The Secret Garden of Neuronal circRNAs.
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Gasparini S, Licursi V, Presutti C, and Mannironi C
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- Animals, Brain growth & development, Brain metabolism, Humans, Mental Disorders genetics, Mental Disorders metabolism, MicroRNAs genetics, MicroRNAs metabolism, Transcriptome, Gene Expression Regulation, Gene Regulatory Networks, Neurons metabolism, RNA, Circular genetics, RNA, Circular metabolism
- Abstract
High-throughput transcriptomic profiling approaches have revealed that circular RNAs (circRNAs) are important transcriptional gene products, identified across a broad range of organisms throughout the eukaryotic tree of life. In the nervous system, they are particularly abundant, developmentally regulated, region-specific, and enriched in genes for neuronal proteins and synaptic factors. These features suggested that circRNAs are key components of an important layer of neuronal gene expression regulation, with known and anticipated functions. Here, we review major recognized aspects of circRNA biogenesis, metabolism and biological activities, examining potential new functions in the context of the nervous system.
- Published
- 2020
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17. Differential Expression of Hippocampal Circular RNAs in the BTBR Mouse Model for Autism Spectrum Disorder.
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Gasparini S, Del Vecchio G, Gioiosa S, Flati T, Castrignano T, Legnini I, Licursi V, Ricceri L, Scattoni ML, Rinaldi A, Presutti C, and Mannironi C
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- Animals, Autism Spectrum Disorder genetics, Brain Chemistry, Gene Expression Profiling, Gene Ontology, Humans, Male, Mice, Inbred C57BL, Mice, Inbred Strains genetics, Mice, Mutant Strains genetics, Reverse Transcriptase Polymerase Chain Reaction, Species Specificity, Autism Spectrum Disorder metabolism, Disease Models, Animal, Hippocampus metabolism, Mice, Inbred Strains metabolism, Mice, Mutant Strains metabolism, RNA, Circular biosynthesis, RNA, Messenger biosynthesis
- Abstract
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition with unknown etiology. Recent experimental evidences suggest the contribution of non-coding RNAs (ncRNAs) in the pathophysiology of ASD. In this work, we aimed to investigate the expression profile of the ncRNA class of circular RNAs (circRNAs) in the hippocampus of the BTBR T + tf/J (BTBR) mouse model and age-matched C57BL/6J (B6) mice. Alongside, we analyzed BTBR hippocampal gene expression profile to evaluate possible correlations between the differential abundance of circular and linear gene products. From RNA sequencing data, we identified circRNAs highly modulated in BTBR mice. Thirteen circRNAs and their corresponding linear isoforms were validated by RT-qPCR analysis. The BTBR-regulated circCdh9 was better characterized in terms of molecular structure and expression, highlighting altered levels not only in the hippocampus, but also in the cerebellum, prefrontal cortex, and amygdala. Finally, gene expression analysis of the BTBR hippocampus pinpointed altered biological and molecular pathways relevant for the ASD phenotype. By comparison of circRNA and gene expression profiles, we identified 6 genes significantly regulated at either circRNA or mRNA gene products, suggesting low overall correlation between circRNA and host gene expression. In conclusion, our results indicate a consistent deregulation of circRNA expression in the hippocampus of BTBR mice. ASD-related circRNAs should be considered in functional studies to identify their contribution to the etiology of the disorder. In addition, as abundant and highly stable molecules, circRNAs represent interesting potential biomarkers for autism.
- Published
- 2020
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18. JARID1B expression and its function in DNA damage repair are tightly regulated by miRNAs in breast cancer.
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Mocavini I, Pippa S, Licursi V, Paci P, Trisciuoglio D, Mannironi C, Presutti C, and Negri R
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- Biomarkers, Tumor, Breast Neoplasms metabolism, Breast Neoplasms mortality, Breast Neoplasms pathology, Cell Cycle genetics, Cell Line, Tumor, Epigenesis, Genetic, Female, Gene Expression Profiling, Genes, Reporter, Humans, RNA Interference, Radiation Tolerance genetics, Reproducibility of Results, Transcriptome, Breast Neoplasms genetics, DNA Damage, DNA Repair, Gene Expression Regulation, Neoplastic, Jumonji Domain-Containing Histone Demethylases genetics, MicroRNAs genetics, Nuclear Proteins genetics, Repressor Proteins genetics
- Abstract
JARID1B/KDM5B histone demethylase's mRNA is markedly overexpressed in breast cancer tissues and cell lines and the protein has been shown to have a prominent role in cancer cell proliferation and DNA repair. However, the mechanism of its post-transcriptional regulation in cancer cells remains elusive. We performed a computational analysis of transcriptomic data from a set of 103 breast cancer patients, which, along with JARID1B upregulation, showed a strong downregulation of 2 microRNAs (miRNAs), mir-381 and mir-486, potentially targeting its mRNA. We showed that both miRNAs can target JARID1B 3'UTR and reduce luciferase's activity in a complementarity-driven repression assay. Moreover, MCF7 breast cancer cells overexpressing JARID1B showed a strong protein reduction when transfected with mir-486. This protein's decrease is accompanied by accumulation of DNA damage, enhanced radiosensitivity and increase of BRCA1 mRNA, 3 features previously correlated with JARID1B silencing. These results enlighten an important role of a miRNA's circuit in regulating JARID1B's activity and suggest new perspectives for epigenetic therapies., (© 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)
- Published
- 2019
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19. miR-135a Regulates Synaptic Transmission and Anxiety-Like Behavior in Amygdala.
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Mannironi C, Biundo A, Rajendran S, De Vito F, Saba L, Caioli S, Zona C, Ciotti T, Caristi S, Perlas E, Del Vecchio G, Bozzoni I, Rinaldi A, Mele A, and Presutti C
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- Adaptor Proteins, Vesicular Transport genetics, Adaptor Proteins, Vesicular Transport metabolism, Amygdala pathology, Animals, Cell Line, Tumor, Excitatory Postsynaptic Potentials, Gene Expression Regulation, Gene Knockdown Techniques, Hippocampus pathology, Humans, Mice, Inbred C57BL, MicroRNAs genetics, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Neurons metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Stress, Physiological genetics, Amygdala metabolism, Amygdala physiopathology, Anxiety genetics, Anxiety physiopathology, Behavior, Animal, MicroRNAs metabolism, Synaptic Transmission genetics
- Abstract
MicroRNAs are a class of non-coding RNAs with a growing relevance in the regulation of gene expression related to brain function and plasticity. They have the potential to orchestrate complex phenomena, such as the neuronal response to homeostatic challenges. We previously demonstrated the involvement of miR-135a in the regulation of early stress response. In the present study, we examine the role of miR-135a in stress-related behavior. We show that the knockdown (KD) of miR-135a in the mouse amygdala induces an increase in anxiety-like behavior. Consistently with behavioral studies, electrophysiological experiments in acute brain slices indicate an increase of amygdala spontaneous excitatory postsynaptic currents, as a result of miR-135a KD. Furthermore, we presented direct evidences, by in vitro assays and in vivo miRNA overexpression in the amygdala, that two key regulators of synaptic vesicle fusion, complexin-1 and complexin-2, are direct targets of miR-135a. In vitro analysis of miniature excitatory postsynaptic currents on miR-135a KD primary neurons indicates unpaired quantal excitatory neurotransmission. Finally, increased levels of complexin-1 and complexin-2 proteins were detected in the mouse amygdala after acute stress, accordingly to the previously observed stress-induced miR-135a downregulation. Overall, our results unravel a previously unknown miRNA-dependent mechanism in the amygdala for regulating anxiety-like behavior, providing evidences of a physiological role of miR-135a in the modulation of presynaptic mechanisms of glutamatergic neurotransmission.
- Published
- 2018
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20. Leptin induction following irradiation is a conserved feature in mammalian epithelial cells and tissues.
- Author
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Licursi V, Cestelli Guidi M, Del Vecchio G, Mannironi C, Presutti C, Amendola R, and Negri R
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- Animals, Cells, Cultured, Dose-Response Relationship, Drug, Humans, Mice, Radiation Dosage, Epithelial Cells metabolism, Epithelial Cells radiation effects, Leptin biosynthesis, Transcriptome physiology, Transcriptome radiation effects
- Abstract
Purpose: Leptin (LEP) is a peptide hormone with multiple physiological functions. Besides its systemic actions, it has important peripheral roles such as a mitogen action on keratinocytes following skin lesions. We previously showed that LEP mRNA is significantly induced in response to neutron irradiation in mouse skin and that the protein increases in the irradiated epidermis and in the related subcutaneous adipose tissue. In this work, we investigated the post-transcriptional regulation of LEP by miRNAs and the conservation of LEP's role in radiation response in human cells., Methods: We used microarray analysis and real-time polymerase chain reaction (RT-PCR) to analyze modulation of miRNAs potentially targeting LEP in mouse skin following irradiation and bioinformatic analysis of transcriptome of irradiated human cell lines and cancer tissues from radiotherapy-treated patients to evaluate LEP expression., Results and Conclusions: We show that a network of miRNAs potentially targeting LEP mRNA is modulated in irradiated mouse skin and that LEP itself is significantly modulated by irradiation in human epithelial cell lines and in breast cancer tissues from radiotherapy-treated patients. These results confirm and extend the previous evidence that LEP has a general and important role in the response of mammalian cells to irradiation.
- Published
- 2017
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21. Antitumor Effects of Epidrug/IFNα Combination Driven by Modulated Gene Signatures in Both Colorectal Cancer and Dendritic Cells.
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Fragale A, Romagnoli G, Licursi V, Buoncervello M, Del Vecchio G, Giuliani C, Parlato S, Leone C, De Angelis M, Canini I, Toschi E, Belardelli F, Negri R, Capone I, Presutti C, and Gabriele L
- Subjects
- Antineoplastic Combined Chemotherapy Protocols administration & dosage, Antineoplastic Combined Chemotherapy Protocols immunology, Azacitidine administration & dosage, Carcinogenesis immunology, Cell Line, Tumor, Colorectal Neoplasms immunology, Colorectal Neoplasms pathology, Dendritic Cells immunology, Depsipeptides administration & dosage, Gene Expression Regulation, Neoplastic drug effects, Gene Expression Regulation, Neoplastic immunology, Humans, Interferon Regulatory Factor-7 genetics, Interferon Regulatory Factors genetics, Interferon-alpha genetics, Interferon-gamma genetics, Nitric Oxide Synthase Type II genetics, Receptors, Cytokine genetics, Receptors, Interferon, Signal Transduction drug effects, Carcinogenesis drug effects, Colorectal Neoplasms drug therapy, Dendritic Cells drug effects, Interferon-alpha immunology
- Abstract
Colorectal cancer results from the progressive accumulation of genetic and epigenetic alterations. IFN signaling defects play an important role in the carcinogenesis process, in which the inability of IFN transcription regulatory factors (IRF) to access regulatory sequences in IFN-stimulated genes (ISG) in tumors and in immune cells may be pivotal. We reported that low-dose combination of two FDA-approved epidrugs, azacytidine (A) and romidepsin (R), with IFNα2 (ARI) hampers the aggressiveness of both colorectal cancer metastatic and stem cells in vivo and triggers immunogenic cell death signals that stimulate dendritic cell (DC) function. Here, we investigated the molecular signals induced by ARI treatment and found that this drug combination increased the accessibility to regulatory sequences of ISGs and IRFs that were epigenetically silenced in both colorectal cancer cells and DCs. Likewise, specific ARI-induced histone methylation and acetylation changes marked epigenetically affected ISG promoters in both metastatic cancer cells and DCs. Analysis by ChIP-seq confirmed such ARI-induced epigenetically regulated IFN signature. The activation of this signal endowed DCs with a marked migratory capability. Our results establish a direct correlation between reexpression of silenced ISGs by epigenetic control and ARI anticancer activity and provide new knowledge for the development of innovative combined therapeutic strategies for colorectal cancer. Cancer Immunol Res; 5(7); 604-16. ©2017 AACR ., (©2017 American Association for Cancer Research.)
- Published
- 2017
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22. miR-142-3p Is a Key Regulator of IL-1β-Dependent Synaptopathy in Neuroinflammation.
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Mandolesi G, De Vito F, Musella A, Gentile A, Bullitta S, Fresegna D, Sepman H, Di Sanza C, Haji N, Mori F, Buttari F, Perlas E, Ciotti MT, Hornstein E, Bozzoni I, Presutti C, and Centonze D
- Subjects
- Adult, Animals, Cells, Cultured, Encephalomyelitis, Autoimmune, Experimental pathology, Female, Gene Knock-In Techniques, Humans, Inflammation metabolism, Inflammation pathology, Male, Mice, Mice, Inbred C57BL, MicroRNAs cerebrospinal fluid, Middle Aged, Multiple Sclerosis, Relapsing-Remitting cerebrospinal fluid, Multiple Sclerosis, Relapsing-Remitting diagnosis, Synapses pathology, Encephalomyelitis, Autoimmune, Experimental metabolism, Interleukin-1beta biosynthesis, MicroRNAs biosynthesis, Multiple Sclerosis, Relapsing-Remitting metabolism, Synapses metabolism
- Abstract
MicroRNAs (miRNA) play an important role in post-transcriptional gene regulation of several physiological and pathological processes. In multiple sclerosis (MS), a chronic inflammatory and degenerative disease of the CNS, and in its mouse model, the experimental autoimmune encephalomyelitis (EAE), miRNA dysregulation has been mainly related to immune system dysfunction and white matter (WM) pathology. However, little is known about their role in gray matter pathology. Here, we explored miRNA involvement in the inflammation-driven alterations of synaptic structure and function, collectively known as synaptopathy, a neuropathological process contributing to excitotoxic neurodegeneration in MS/EAE. Particularly, we observed that miR-142-3p is increased in the CSF of patients with active MS and in EAE brains. We propose miR-142-3p as a molecular mediator of the IL-1β-dependent downregulation of the glial glutamate-aspartate transporter (GLAST), which causes an enhancement of the glutamatergic transmission in the EAE cerebellum. The synaptic abnormalities mediated by IL-1β and the clinical and neuropathological manifestations of EAE disappeared in miR-142 knock-out mice. Furthermore, we observed that in vivo miR-142-3p inhibition, either by a preventive and local treatment or by a therapeutic and systemic strategy, abolished IL-1β- and GLAST-dependent synaptopathy in EAE wild-type mice. Consistently, miR-142-3p was responsible for the glutamatergic synaptic alterations caused by CSF of patients with MS, and CSF levels of miR-142-3p correlated with prospective MS disease progression. Our findings highlight miR-142-3p as key molecular player in IL-1β-mediated synaptic dysfunction, possibly leading to excitotoxic damage in both EAE and MS diseases. Inhibition of miR-142-3p could be neuroprotective in MS., Significance Statement: Current studies suggest the role of glutamate excitotoxicity in the development and progression of multiple sclerosis (MS) and of its mouse model experimental autoimmune encephalomyelitis (EAE). The molecular mechanisms linking inflammation and synaptic alterations in MS/EAE are still unknown. Here, we identified miR-142-3p as a determinant molecular actor in inflammation-dependent synaptopathy typical of both MS and EAE. miR-142-3p was upregulated in the CSF of MS patients and in EAE cerebellum. Inhibition of miR-142-3p, locally in EAE brain and in a MS chimeric ex vivo model, recovered glutamatergic synaptic enhancement typical of EAE/MS. We proved that miR-142-3p promoted the IL-1β-dependent glutamate dysfunction by targeting glutamate-aspartate transporter (GLAST), a crucial glial transporter involved in glutamate homeostasis. Finally, we suggest miR-142-3p as a negative prognostic factor in patients with relapsing-remitting multiple sclerosis., (Copyright © 2017 the authors 0270-6474/17/370547-16$15.00/0.)
- Published
- 2017
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23. RNA-binding protein HuR and the members of the miR-200 family play an unconventional role in the regulation of c-Jun mRNA.
- Author
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Del Vecchio G, De Vito F, Saunders SJ, Risi A, Mannironi C, Bozzoni I, and Presutti C
- Subjects
- 3' Untranslated Regions, Binding Sites, ELAV-Like Protein 1 genetics, HEK293 Cells, HeLa Cells, Humans, JNK Mitogen-Activated Protein Kinases metabolism, MCF-7 Cells, MicroRNAs metabolism, Protein Binding, RNA Stability, RNA, Messenger genetics, RNA, Messenger metabolism, ELAV-Like Protein 1 metabolism, JNK Mitogen-Activated Protein Kinases genetics, MicroRNAs genetics
- Abstract
Post-transcriptional gene regulation is a fundamental step for coordinating cellular response in a variety of processes. RNA-binding proteins (RBPs) and microRNAs (miRNAs) are the most important factors responsible for this regulation. Here we report that different components of the miR-200 family are involved in c-Jun mRNA regulation with the opposite effect. While miR-200b inhibits c-Jun protein production, miR-200a tends to increase the JUN amount through a stabilization of its mRNA. This action is dependent on the presence of the RBP HuR that binds the 3'UTR of c-Jun mRNA in a region including the mir-200a binding site. The position of the binding site is fundamental; by mutating this site, we demonstrate that the effect is not micro-RNA specific. These results indicate that miR-200a triggers a microRNA-mediated stabilization of c-Jun mRNA, promoting the binding of HuR with c-Jun mRNA. This is the first example of a positive regulation exerted by a microRNA on an important oncogene in proliferating cells., (© 2016 Del Vecchio et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
- Published
- 2016
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24. Modulation of microRNA editing, expression and processing by ADAR2 deaminase in glioblastoma.
- Author
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Tomaselli S, Galeano F, Alon S, Raho S, Galardi S, Polito VA, Presutti C, Vincenti S, Eisenberg E, Locatelli F, and Gallo A
- Subjects
- Adolescent, Animals, Brain enzymology, Brain Neoplasms enzymology, Brain Neoplasms pathology, Cell Line, Tumor, Cell Movement, Cell Proliferation, Down-Regulation genetics, Gene Expression Profiling, Gene Silencing, Glioblastoma pathology, HEK293 Cells, Humans, Mice, MicroRNAs metabolism, Models, Biological, Adenosine Deaminase metabolism, Brain Neoplasms genetics, Gene Expression Regulation, Neoplastic, Glioblastoma enzymology, Glioblastoma genetics, MicroRNAs genetics, RNA Editing genetics, RNA-Binding Proteins metabolism
- Abstract
Background: ADAR enzymes convert adenosines to inosines within double-stranded RNAs, including microRNA (miRNA) precursors, with important consequences on miRNA retargeting and expression. ADAR2 activity is impaired in glioblastoma and its rescue has anti-tumoral effects. However, how ADAR2 activity may impact the miRNome and the progression of glioblastoma is not known., Results: By integrating deep-sequencing and array approaches with bioinformatics analyses and molecular studies, we show that ADAR2 is essential to edit a small number of mature miRNAs and to significantly modulate the expression of about 90 miRNAs in glioblastoma cells. Specifically, the rescue of ADAR2 activity in cancer cells recovers the edited miRNA population lost in glioblastoma cell lines and tissues, and rebalances expression of onco-miRNAs and tumor suppressor miRNAs to the levels observed in normal human brain. We report that the major effect of ADAR2 is to reduce the expression of a large number of miRNAs, most of which act as onco-miRNAs. ADAR2 can edit miR-222/221 and miR-21 precursors and decrease the expression of the corresponding mature onco-miRNAs in vivo and in vitro, with important effects on cell proliferation and migration., Conclusions: Our findings disclose an additional layer of complexity in miRNome regulation and provide information to better understand the impact of ADAR2 editing enzyme in glioblastoma. We propose that ADAR2 is a key factor for maintaining edited-miRNA population and balancing the expression of several essential miRNAs involved in cancer.
- Published
- 2015
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25. Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.
- Author
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Alteri A, De Vito F, Messina G, Pompili M, Calconi A, Visca P, Mottolese M, Presutti C, and Grossi M
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- Adenocarcinoma genetics, Adenocarcinoma metabolism, Cell Line, Transformed, Cell Line, Tumor, Cell Transformation, Neoplastic pathology, Cyclin D1 metabolism, Humans, Lung Neoplasms genetics, Lung Neoplasms metabolism, MicroRNAs genetics, Muscle, Skeletal cytology, Muscle, Skeletal metabolism, Organ Specificity, Cell Differentiation genetics, Cell Transformation, Neoplastic genetics, Cyclin D1 genetics, MicroRNAs metabolism
- Abstract
miR-206, a member of the so-called myomiR family, is largely acknowledged as a specific, positive regulator of skeletal muscle differentiation. A growing body of evidence also suggests a tumor suppressor function for miR-206, as it is frequently downregulated in various types of cancers. In this study, we show that miR-206 directly targets cyclin D1 and contributes to the regulation of CCND1 gene expression in both myogenic and non-muscle, transformed cells. We demonstrate that miR-206, either exogenous or endogenous, reduces cyclin D1 levels and proliferation rate in C2C12 cells without promoting differentiation, and that miR-206 knockdown in terminally differentiated C2C12 cells leads to cyclin D1 accumulation in myotubes, indicating that miR-206 might be involved in the maintenance of the post-mitotic state. Targeting of cyclin D1 might also account, at least in part, for the tumor-suppressor activity suggested for miR-206 in previous studies. Accordingly, the analysis of neoplastic and matched normal lung tissues reveals that miR-206 downregulation in lung tumors correlates, in most cases, with higher cyclin D1 levels. Moreover, gain-of-function experiments with cancer-derived cell lines and with in vitro transformed cells indicate that miR-206-mediated cyclin D1 repression is directly coupled to growth inhibition. Altogether, our data highlight a novel activity for miR-206 in skeletal muscle differentiation and identify cyclin D1 as a major target that further strengthens the tumor suppressor function proposed for miR-206.
- Published
- 2013
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26. Acute stress alters amygdala microRNA miR-135a and miR-124 expression: inferences for corticosteroid dependent stress response.
- Author
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Mannironi C, Camon J, De Vito F, Biundo A, De Stefano ME, Persiconi I, Bozzoni I, Fragapane P, Mele A, and Presutti C
- Subjects
- Animals, Base Sequence, Male, Mice, Mice, Inbred C57BL, Receptors, Mineralocorticoid genetics, Receptors, Mineralocorticoid metabolism, Adrenal Cortex Hormones metabolism, Amygdala metabolism, Gene Expression Regulation, MicroRNAs genetics, Stress, Psychological genetics, Stress, Psychological metabolism
- Abstract
The amygdala is a brain structure considered a key node for the regulation of neuroendocrine stress response. Stress-induced response in amygdala is accomplished through neurotransmitter activation and an alteration of gene expression. MicroRNAs (miRNAs) are important regulators of gene expression in the nervous system and are very well suited effectors of stress response for their ability to reversibly silence specific mRNAs. In order to study how acute stress affects miRNAs expression in amygdala we analyzed the miRNA profile after two hours of mouse restraint, by microarray analysis and reverse transcription real time PCR. We found that miR-135a and miR-124 were negatively regulated. Among in silico predicted targets we identified the mineralocorticoid receptor (MR) as a target of both miR-135a and miR-124. Luciferase experiments and endogenous protein expression analysis upon miRNA upregulation and inhibition allowed us to demonstrate that mir-135a and mir-124 are able to negatively affect the expression of the MR. The increased levels of the amygdala MR protein after two hours of restraint, that we analyzed by western blot, negatively correlate with miR-135a and miR-124 expression. These findings point to a role of miR-135a and miR-124 in acute stress as regulators of the MR, an important effector of early stress response.
- Published
- 2013
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27. IFN-α regulates Blimp-1 expression via miR-23a and miR-125b in both monocytes-derived DC and pDC.
- Author
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Parlato S, Bruni R, Fragapane P, Salerno D, Marcantonio C, Borghi P, Tataseo P, Ciccaglione AR, Presutti C, Romagnoli G, Bozzoni I, Belardelli F, and Gabriele L
- Subjects
- Cell Differentiation drug effects, Cell Differentiation genetics, Dendritic Cells drug effects, Gene Expression Profiling, HeLa Cells, Humans, Phenotype, Positive Regulatory Domain I-Binding Factor 1, Repressor Proteins metabolism, Dendritic Cells cytology, Dendritic Cells metabolism, Interferon-alpha pharmacology, MicroRNAs genetics, Monocytes cytology, Repressor Proteins genetics
- Abstract
Type I interferon (IFN-I) have emerged as crucial mediators of cellular signals controlling DC differentiation and function. Human DC differentiated from monocytes in the presence of IFN-α (IFN-α DC) show a partially mature phenotype and a special capability of stimulating CD4+ T cell and cross-priming CD8+ T cells. Likewise, plasmacytoid DC (pDC) are blood DC highly specialized in the production of IFN-α in response to viruses and other danger signals, whose functional features may be shaped by IFN-I. Here, we investigated the molecular mechanisms stimulated by IFN-α in driving human monocyte-derived DC differentiation and performed parallel studies on peripheral unstimulated and IFN-α-treated pDC. A specific miRNA signature was induced in IFN-α DC and selected miRNAs, among which miR-23a and miR-125b, proved to be negatively associated with up-modulation of Blimp-1 occurring during IFN-α-driven DC differentiation. Of note, monocyte-derived IFN-α DC and in vitro IFN-α-treated pDC shared a restricted pattern of miRNAs regulating Blimp-1 expression as well as some similar phenotypic, molecular and functional hallmarks, supporting the existence of a potential relationship between these DC populations. On the whole, these data uncover a new role of Blimp-1 in human DC differentiation driven by IFN-α and identify Blimp-1 as an IFN-α-mediated key regulator potentially accounting for shared functional features between IFN-α DC and pDC.
- Published
- 2013
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28. HUVEC respond to radiation by inducing the expression of pro-angiogenic microRNAs.
- Author
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Vincenti S, Brillante N, Lanza V, Bozzoni I, Presutti C, Chiani F, Etna MP, and Negri R
- Subjects
- 3' Untranslated Regions genetics, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, Binding Sites, Capillaries cytology, Cell Nucleus metabolism, Cell Nucleus radiation effects, Dose-Response Relationship, Radiation, Gene Expression Regulation genetics, HeLa Cells, Humans, Linear Energy Transfer, Oxidative Stress genetics, Oxidative Stress radiation effects, Proto-Oncogene Proteins c-myc metabolism, X-Rays, Endothelial Cells metabolism, Endothelial Cells radiation effects, Gene Expression Regulation radiation effects, MicroRNAs genetics, Neovascularization, Physiologic genetics, Neovascularization, Physiologic radiation effects, Umbilical Cord cytology
- Abstract
MicroRNAs (miRNAs) represent a class of small non-coding RNAs that control gene expression by targeting mRNAs and triggering either repression of translation or RNA degradation. They have been shown to be involved in a variety of biological processes such as development, differentiation and cell cycle control, but little is known about their involvement in the response to irradiation. We showed here that in human umbilical vein endothelial cells (HUVEC) some miRNAs previously shown to have a crucial role in vascular biology are transiently modulated in response to a clinically relevant dose of ionizing radiation. In particular we identified an early transcriptional induction of several members of the microRNA cluster 17-92 and other microRNAs already known to be related to angiogenesis. At the same time we observed a peculiar behavior of the miR-221/222 cluster, suggesting an important role of these microRNAs in HUVEC homeostasis. We observed an increased efficiency in the formation of capillary-like structures in irradiated HUVEC. These results could lead to a new interpretation of the effect of ionizing radiation on endothelial cells and on the response of tumor endothelial bed cells to radiotherapy.
- Published
- 2011
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29. Smad-interacting protein-1 and microRNA 200 family define a nitric oxide-dependent molecular circuitry involved in embryonic stem cell mesendoderm differentiation.
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Rosati J, Spallotta F, Nanni S, Grasselli A, Antonini A, Vincenti S, Presutti C, Colussi C, D'Angelo C, Biroccio A, Farsetti A, Capogrossi MC, Illi B, and Gaetano C
- Subjects
- Animals, Cells, Cultured, Chromatin Assembly and Disassembly, Embryonic Stem Cells drug effects, Gene Expression Regulation, Developmental, Homeodomain Proteins genetics, Leukemia Inhibitory Factor metabolism, Mice, Nitric Oxide Donors pharmacology, RNA Interference, RNA, Messenger metabolism, Repressor Proteins genetics, Telomerase metabolism, Time Factors, Transcription, Genetic, Transfection, Zinc Finger E-box Binding Homeobox 2, Cell Differentiation drug effects, Embryonic Stem Cells metabolism, Homeodomain Proteins metabolism, MicroRNAs metabolism, Nitric Oxide metabolism, Repressor Proteins metabolism, Signal Transduction drug effects
- Abstract
Objective: Smad-interacting protein-1 (Sip1/ZEB2) is a transcriptional repressor of the telomerase reverse transcriptase catalytic subunit (Tert) and has recently been identified as a key regulator of embryonic cell fate with a phenotypic effect similar, in our opinion, to that reported for nitric oxide (NO). Remarkably, SIP1/ZEB2 is a known target of the microRNA 200 (miR-200) family. In this light, we postulated that Sip1/ZEB2 and the miR-200 family could play a role during the NO-dependent differentiation of mES., Methods and Results: The results of the present study show that Sip1/ZEB2 expression is downregulated during the NO-dependent expression of mesendoderm and early cardiovascular precursor markers, including Flk1 and CXCR4 in mES. Coincidently, members of the miR-200 family, namely miR-429, -200a, -200b, and -200c, were transcriptionally induced in parallel to mouse Tert. This regulation occurred at the level of chromatin. Remarkably, miR-429/miR-200a overexpression or Sip1/ZEB2 knockdown by short hairpin RNA interference elicited a gene expression pattern similar to that of NO regardless of the presence of leukemia inhibitory factor., Conclusions: These results are the first demonstrating that the miR-200 family and Sip1/ZEB2 transcription factor are regulated by NO, indicating an unprecedented molecular circuitry important for telomerase regulation and early differentiation of mES.
- Published
- 2011
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30. Stress induces region specific alterations in microRNAs expression in mice.
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Rinaldi A, Vincenti S, De Vito F, Bozzoni I, Oliverio A, Presutti C, Fragapane P, and Mele A
- Subjects
- Animals, Gene Expression Profiling methods, Hippocampus metabolism, Male, Mice, MicroRNAs classification, MicroRNAs genetics, Oligonucleotide Array Sequence Analysis methods, Stress, Psychological pathology, Time Factors, Gene Expression Regulation physiology, MicroRNAs metabolism, Stress, Psychological metabolism
- Abstract
Several studies have demonstrated that exposure to both acute and chronic aversive stimuli can affect neural activity in different brain areas. In particular it has been shown that stressful events can induce not only short-term changes in neural transmission and gene regulation, but also long-term changes that can lead to structural modification. In this study we investigated, in CD1 mice, the effects of single or repeated exposures to restraint stress (2h for 1 or 5 consecutive days) in the frontal cortex on a crucial class of gene expression regulators, the microRNAs (miRs).First we performed a microarray profiling on RNA extracted from the frontal cortex of mice exposed to acute or repeated restraint stress. The results indicated a prominent increase in the expression levels of different miRs after acute stress while only minor changes were observed after repeated restraint. The Northern blot analysis on selected miRs confirmed an increase after acute restraint for let-7a, miR-9 and miR 26-a/b. Finally, Northern blot analysis of the selected miRs on RNA extracted from the hippocampus of stressed mice demonstrated that such changes were region specific, as no differences were observed in the hippocampus. These data suggest that control of mRNA translation through miRs is an additional mechanism by which stressful events regulates protein expression in the frontal cortex., (Copyright 2009 Elsevier B.V. All rights reserved.)
- Published
- 2010
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31. Role of Hog1 and Yaf9 in the transcriptional response of Saccharomyces cerevisiae to cesium chloride.
- Author
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Del Vescovo V, Casagrande V, Bianchi MM, Piccinni E, Frontali L, Militti C, Fardeau V, Devaux F, Di Sanza C, Presutti C, and Negri R
- Subjects
- Acetyltransferases genetics, Adaptation, Physiological drug effects, Adaptation, Physiological genetics, Cell Wall drug effects, Cell Wall genetics, Cluster Analysis, Dose-Response Relationship, Drug, Gene Expression Profiling, Gene Expression Regulation, Fungal drug effects, Histone Acetyltransferases, Metals, Alkali pharmacology, Mitogen-Activated Protein Kinases genetics, Mitogen-Activated Protein Kinases metabolism, Oligonucleotide Array Sequence Analysis, Organisms, Genetically Modified, Osmolar Concentration, Phosphorylation drug effects, Saccharomyces cerevisiae drug effects, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism, Signal Transduction drug effects, Signal Transduction genetics, Acetyltransferases physiology, Cesium pharmacology, Chlorides pharmacology, Mitogen-Activated Protein Kinases physiology, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins physiology, Transcription, Genetic drug effects
- Abstract
We analyzed the global transcriptional response of Saccharomyces cerevisiae cells exposed to different concentrations of CsCl in the growth medium and at different times after addition. Early responsive genes were mainly involved in cell wall structure and biosynthesis. About half of the induced genes were previously shown to respond to other alkali metal cations in a Hog1-dependent fashion. Western blot analysis confirmed that cesium concentrations as low as 100 mM activate Hog1 phosphorylation. Another important fraction of the cesium-modulated genes requires Yaf9p for full responsiveness as shown by the transcriptome of a yaf9-deleted strain in the presence of cesium. We showed that a cell wall-restructuring process promptly occurs in response to cesium addition, which is dependent on the presence of both Hog1 and Yaf9 proteins. Moreover, the sensitivity to low concentration of cesium of the yaf9-deleted strain is not observed in a strain carrying the hog1/yaf9 double deletion. We conclude that the observed early transcriptional modulation of cell wall genes has a crucial role in S. cerevisiae adaptation to cesium.
- Published
- 2008
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32. The position of yeast snoRNA-coding regions within host introns is essential for their biosynthesis and for efficient splicing of the host pre-mRNA.
- Author
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Vincenti S, De Chiara V, Bozzoni I, and Presutti C
- Subjects
- Mutation, Nuclear Proteins metabolism, RNA Precursors genetics, RNA, Fungal genetics, RNA, Small Nucleolar genetics, Ribonucleoproteins, Small Nucleolar metabolism, Saccharomyces cerevisiae Proteins metabolism, Transcription, Genetic, Introns genetics, RNA Precursors metabolism, RNA Splicing, RNA, Fungal metabolism, RNA, Small Nucleolar biosynthesis, Saccharomyces cerevisiae genetics
- Abstract
Genomic location of sequences encoding small nucleolar RNAs (snoRNAs) is peculiar in all eukaryotes from yeast to mammals: most of them are encoded within the introns of host genes. In Saccharomyces cerevisiae, seven snoRNAs show this location. In this work we demonstrate that the position of snoRNA-coding regions with respect to splicing consensus sequences is critical: yeast strains expressing mutant constructs containing shorter or longer spacers (the regions between snoRNA ends and intron splice sites) show a drop in accumulation of U24 and U18 snoRNAs. Further mutational analysis demonstrates that altering the distance between the 3' end of the snoRNA and the branch point is the most important constraint for snoRNA biosynthesis, and that stable external stems, which are sometimes present in introns containing snoRNAs, can overcome the positional effect. Surprisingly enough, splicing of the host introns is clearly affected in most of these constructs indicating that, at least in S. cerevisiae, an incorrect location of snoRNA-coding sequences within the host intron is detrimental to the splicing process. This is different with respect to what was demonstrated in mammals, where the activity of the splicing machinery seems to be dominant with respect to the assembly of snoRNPs, and it is not affected by the location of snoRNA sequences. We also show that intronic box C/D snoRNA recognition and assembly of snoRNPs occur during transcription when splicing sequences are recognized.
- Published
- 2007
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33. Non coding RNA and brain.
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Presutti C, Rosati J, Vincenti S, and Nasi S
- Subjects
- Animals, Gene Expression Regulation genetics, Humans, Models, Biological, RNA, Untranslated genetics, Brain metabolism, RNA, Untranslated metabolism
- Abstract
Small non coding RNAs are a group of very different RNA molecules, present in virtually all cells, with a wide spectrum of regulatory functions which include RNA modification and regulation of protein synthesis. They have been isolated and characterized in all organisms and tissues, from Archaeobacteria to mammals. In mammalian brain there are a number of these small molecules, which are involved in neuronal differentiation as well as, possibly, in learning and memory. In this manuscript, we analyze the present knowledge about the function of the most important groups of small non-coding RNA present in brain: small nucleolar RNAs, small cytoplasmic RNAs, and microRNAs. The last ones, in particular, appear to be critical for dictating neuronal cell identity during development and to play an important role in neurite growth, synaptic development and neuronal plasticity.
- Published
- 2006
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34. Separation and characterisation of sphingoceramides by high-performance liquid chromatography-electrospray ionisation mass spectrometry.
- Author
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Camera E, Picardo M, Presutti C, Catarcini P, and Fanali S
- Subjects
- Chromatography, Chromatography, Liquid methods, Epidermal Cells, Humans, Ions, Keratinocytes metabolism, Lipids analysis, Lipids chemistry, Mass Spectrometry methods, Skin cytology, Skin metabolism, Sphingosine chemistry, Time Factors, Ceramides chemistry, Chromatography, High Pressure Liquid methods, Spectrometry, Mass, Electrospray Ionization methods
- Abstract
We developed a simple and reliable analytical method for the quantification and the characterization of ceramides extracted from biological samples by high-performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (ESI/MS/MS). The chromatographic separation of analytes was carried out in a RP8 column, eluting with a methanol-water mixture in gradient elution mode. The separated lipids were detected by total ion monitoring and characterised by MS/MS spectra; quantitative analysis was performed by integrating the extracted ion peaks obtained in the negative ion mode. Good repeatability was obtained for retention time (0.3-2%), peak area ratio (A(S)/A(IS), 2-8%), as well as limit of detection (LOD, 5-26 pg) and quantification (LOQ, 13-53 pg). The method was validated for the analysis of N-palmitoyl-D-erythro-sphingosine (Cer16), N-stearoyl-D-erythro-sphingosine (Cer18), N-tetracosanoyl-D-erythro-sphingosine (N24:0, lignoceric ceramide, Cer24:0), and N-tetracos-15'-enoyl-D-erythro-sphingosine (N24:1, nervonic ceramide, Cer24:1), giving good results. Lipid mixtures, extracted from skin and epidermal cells, were analysed for their content of the studied ceramides.
- Published
- 2004
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35. Experimental assessment of electromigration properties of background electrolytes in capillary zone electrophoresis.
- Author
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Bousková E, Presutti C, Gebauer P, Fanali S, Beckers JL, and Bocek P
- Subjects
- Acetates isolation & purification, Buffers, Histamine isolation & purification, Histidine isolation & purification, Imidazoles isolation & purification, Electrolytes, Electrophoresis, Capillary methods, Ions isolation & purification
- Abstract
Electromigration dispersion (EMD) properties of background electrolytes (BGEs) used in capillary zone electrophoresis (CZE) are of key importance for the success of an analysis. The knowledge of these properties may serve well for the prediction of the asymmetry of peaks of analytes, for the prediction of unsafe regions where a strong interference of system zones may be expected, and for the selection of optimum conditions where the analytes of interest may give sharp and practically symmetric peaks. Present theories enable one to calculate and predict EMD properties of many BGEs but there is also a lot of BGEs that are beyond the present theoretical models as far as their composition and equilibria involved are considered. This contribution brings a method for assessment of EMD properties of any BGE from easily accessible experimental data. The method proposed is illustrated by model examples both for cationic and anionic separations. Imidazole acetate, histamine acetate, and histidine acetate served as model BGEs for cationic separations; as the model BGE for anionic separations, Tris-borate and sodium-borate BGEs have been selected since these buffers are frequently used and borate is well-known for its complexing equilibria in aqueous solutions.
- Published
- 2004
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36. Evaluation of teicoplanin chiral stationary phases of 3.5 and 5 microm inside diameter silica microparticles by polar-organic mode capillary electrochromatography.
- Author
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Catarcini P, Fanali S, Presutti C, D'Acquarica I, and Gasparrini F
- Subjects
- Acetates chemistry, Acetonitriles chemistry, Adrenergic beta-Antagonists chemistry, Methanol chemistry, Particle Size, Reproducibility of Results, Stereoisomerism, Adrenergic beta-Antagonists analysis, Chromatography, Liquid methods, Electrophoresis, Capillary methods, Silicon Dioxide chemistry, Teicoplanin chemistry
- Abstract
Different types of fused-silica capillaries of 75 microm inside diameter (ID) were packed, namely type A and B, and evaluated for the direct resolution of racemates of several basic compounds by enantioselective capillary electrochromatography (e-CEC). Type A was packed with a chiral stationary phase (CSP) containing teicoplanin (TE) mixed with silica microparticles (3:1 w/w) while type B contained only the TE-CSP. In both cases, particles of different sizes (3.5 and 5 microm ID) were employed. A polar-organic mobile phase containing methanol-acetonitrile (60-40% v/v and 0.05% w/v ammonium acetate was used. Several beta-blockers (alprenolol, oxprenolol, metoprolol, pindolol, salbutamol, propranolol, atenolol, acebutolol) were baseline-enantioresolved with both capillary types, in very short times.
- Published
- 2003
- Full Text
- View/download PDF
37. Enantiomeric separation of acidic compounds of pharmaceutical interest by capillary electrochromatography employing glycopeptide antibiotic stationary phases.
- Author
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Fanali S, Catarcini P, and Presutti C
- Subjects
- Acids, Stereoisomerism, Anti-Bacterial Agents chemistry, Chromatography, Micellar Electrokinetic Capillary methods, Glycopeptides, Pharmaceutical Preparations chemistry
- Abstract
Enantiomeric separation of some selected acidic compounds of pharmaceutical interest belonging to the group of non-steroidal anti-inflammatory drugs were separated by capillary electrochromatography employing silica based glycopeptide antibiotic stationary phases, namely vancomycin or a teicoplanin derivatives (Hepta-Tyr). The vancomycin stationary phase allowed to achieve the chiral resolution of some racemic studied compounds only using mobile phases containing ammonium formate at a relatively low pH 2.5-3.5 and acetonitrile. Employing the teicoplanin derivative stationary phase, good enantiomeric resolution was achieved eluting with mobile phases containing sodium phosphate pH 6-acetonitrile. Enantiomers were moved to the detector because a relatively high reversed electroosmotic flow (due to the positive charge of the stationary phase) and to the electrophoretic mobility of analytes.
- Published
- 2003
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38. Use of short-end injection capillary packed with a glycopeptide antibiotic stationary phase in electrochromatography and capillary liquid chromatography for the enantiomeric separation of hydroxy acids.
- Author
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Fanali S, Catarcini P, Presutti C, Stancanelli R, and Quaglia MG
- Subjects
- Stereoisomerism, Acids isolation & purification, Anti-Bacterial Agents chemistry, Chromatography, High Pressure Liquid instrumentation, Chromatography, Micellar Electrokinetic Capillary instrumentation, Glycopeptides
- Abstract
A new chiral stationary phase (CSP) was prepared by reacting MDL 63,246 (Hepta-Tyr), a glycopeptide antibiotic belonging to the teicoplanin family, with 5-microm diol-silica particles. The CSP mixed with 5-microm amino silica particles (3:1) was packed into 75-microm fused-silica capillaries for only 6.6 cm and used for electrochromatographic experiments analyzing several hydroxy acid enantiomers. A reversed electroosmotic flow carried both analytes and mobile phase towards the anode in a short time (1-3 min), being baseline resolved all the studied analytes. In order to achieve the fastest enantiomeric resolution of the studied hydroxy acids, the effect of several experimental parameters such as mobile phase composition (organic modifier type and concentration, pH of the buffer and ionic strength), capillary temperature and applied voltage on enantioresolution factor, retention time, enantioselectivity were evaluated. The packed capillary column allowed the separation of mandelic acid enantiomers in less than 72 s with resolution factor Rs=2.18 applying a voltage of 30 kV and eluting with a mobile phase composed by 50 mM ammonium acetate (pH 6)-water-acetonitrile (1:4:5, v/v). The CSP was also tested in the capillary liquid chromatography mode resolving all the studied enantiomers applying 12 bar pressure to the mobile phase [50 mM ammonium acetate (pH 6)-water-methanol-acetonitrile, 1:4:2:3, v/v)], however, relatively long analysis times were observed (12-20 min).
- Published
- 2003
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39. A glycopeptide antibiotic chiral stationary phase for the enantiomer resolution of hydroxy acid derivatives by capillary electrochromatography.
- Author
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Fanali S, Catarcini P, Presutti C, Quaglia MG, and Righetti PG
- Subjects
- Acetonitriles, Buffers, Hydrogen-Ion Concentration, Hydroxy Acids analysis, Hydroxy Acids chemistry, Lactates analysis, Mandelic Acids analysis, Molecular Structure, Solvents chemistry, Stereoisomerism, Teicoplanin analogs & derivatives, Teicoplanin chemistry, Temperature, Time, Anti-Bacterial Agents chemistry, Chromatography, Micellar Electrokinetic Capillary methods
- Abstract
Separation of hydroxy acid enantiomers was achieved by using capillary electrochromatography (CEC) employing a chiral stationary phase (CSP) based on MDL 63,246 (Hepta-Tyr), a macrocyclic antibiotic of the teicoplanin family. The chiral selector was chemically bonded to 5 num diol-modified silica particles and the CSP mixed with amino silica (3:1 w/w) was packed into a 75 num ID fused-silica capillary. The CEC experiments were carried out by using an aqueous reversed-phase mode for the enantiomeric resolution of hydroxy acid compounds. Good enantioresolution was achieved for mandelic acid (MA), m-hydroxymandelic acid (m-OH-MA), p-OH-MA, and 3-hydroxy-4-methoxymandelic acid (3-OH-4-MeO-MA). The CEC system was less enantioselective towards 2-phenyllactic acid (2-PhL) and 3-PhL while mandelic acid methyl ester (MA-Et-Est) enantiomers were not resolved. Several experimental parameters, such as organic solvent type and concentration, buffer pH, capillary temperature, on enantioresolution factor, retention time, and retention factor were studied.
- Published
- 2003
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40. Purified box C/D snoRNPs are able to reproduce site-specific 2'-O-methylation of target RNA in vitro.
- Author
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Galardi S, Fatica A, Bachi A, Scaloni A, Presutti C, and Bozzoni I
- Subjects
- Binding Sites, Conserved Sequence, Fungal Proteins chemistry, Macromolecular Substances, Mass Spectrometry, Methylation, Methyltransferases isolation & purification, Nuclear Proteins chemistry, Oligoribonucleotides chemistry, RNA, Ribosomal chemistry, Ribonucleoproteins, Small Nuclear chemistry, Ribonucleoproteins, Small Nucleolar isolation & purification, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins chemistry, Substrate Specificity, Methyltransferases chemistry, RNA, Fungal chemistry, Ribonucleoproteins, Small Nucleolar chemistry
- Abstract
Small nucleolar RNAs (snoRNAs) are associated in ribonucleoprotein particles localized to the nucleolus (snoRNPs). Most of the members of the box C/D family function in directing site-specific 2'-O-methylation of substrate RNAs. Although the selection of the target nucleotide requires the antisense element and the conserved box D or D' of the snoRNA, the methyltransferase activity is supposed to reside in one of the protein components. Through protein tagging of a snoRNP-specific factor, we purified to homogeneity box C/D snoRNPs from the yeast Saccharomyces cerevisiae. Mass spectrometric analysis demonstrated the presence of Nop1p, Nop58p, Nop56p, and Snu13p as integral components of the particle. We show that purified snoRNPs are able to reproduce the site-specific methylation pattern on target RNA and that the predicted S-adenosyl-L-methionine-binding region of Nop1p is responsible for the catalytic activity.
- Published
- 2002
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41. Enantioseparation of amino acid derivatives by capillary zone electrophoresis using vancomycin as chiral selector.
- Author
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Fanali S, Crucianelli M, De Angelis F, and Presutti C
- Subjects
- Anti-Bacterial Agents chemistry, Buffers, Hydrogen-Ion Concentration, Indicators and Reagents chemistry, Stereoisomerism, Temperature, Amino Acids isolation & purification, Electrophoresis, Capillary methods, Vancomycin chemistry
- Abstract
The separation of racemic derivatized amino acids (N-acetyl) into their enantiomers was achieved using capillary zone electrophoresis employing vancomycin as a chiral selector. Due to the strong absorption properties of the chiral selector at the low wavelengths used, the partial-filling countercurrent method was adopted in order to improve method sensitivity. In the separation system studied, the chiral selector filled only a part of the capillary and, due to the appropriate selection of the pH, was moving in the opposite direction of the analytes keeping the detector free from absorbing compounds. The effect of several experimental parameters on the enantioresolution of analytes was studied, e.g., vancomycin concentration (0-5 mM), pH of the background electrolyte (pH 4-7), capillary temperature (15-35 degrees C), and the presence of an organic modifier in the run buffer (methanol or ethanol or n-propanol). N-Acetyl glutamic acid, serine, cystine, tyrosine, and proline were all baseline-resolved into their enantiomers and the enantioresolution factor (R(s)) was increased by raising the vancomycin concentration. pH 4 allowed the baseline resolution of the five studied analytes in the presence of 2.5 mM of chiral selector and an increase in pH caused a decrease of R(s).
- Published
- 2002
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- View/download PDF
42. Enteroinvasive Escherichia coli virulence-plasmid-carried apyrase (apy) and ospB genes are organized as a bicistronic operon and are subject to differential expression.
- Author
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Santapaola D, Casalino M, Petrucca A, Presutti C, Zagaglia C, Berlutti F, Colonna B, and Nicoletti M
- Subjects
- Apyrase biosynthesis, Blotting, Northern, Escherichia coli enzymology, Escherichia coli pathogenicity, Genes, Bacterial, Molecular Sequence Data, Operon, RNA, Messenger biosynthesis, Reverse Transcriptase Polymerase Chain Reaction, Temperature, Transcription, Genetic, Virulence genetics, Apyrase genetics, Escherichia coli genetics, Gene Expression Regulation, Bacterial, Plasmids genetics
- Abstract
In Shigella flexneri and enteroinvasive Escherichia coli (EIEC) the expression of the virulence-plasmid(pINV)-carried potential pathogenesis-associated apy gene, which encodes apyrase (ATP diphosphohydrolase), is regulated by the same regulators that govern the expression of virulence genes. To understand the transcriptional organization of the apy gene, the authors sequenced an 8023 bp PstI fragment of the pINV of EIEC strain HN280, which encompasses apy as well as its adjacent genes. The PstI fragment displays 99% identity with the corresponding fragment of pWR100, the pINV of S. flexneri strain M90T, and contains four genes. One of these genes, ospB, encodes a secreted protein of unknown activity and is located immediately upstream of apy. Analyses of sequence, Northern hybridization, RT-PCR and primer extension data and transcriptional fusions indicated that ospB and apy are co-transcribed as a 2 kb bicistronic, temperature-regulated mRNA from an upstream promoter that precedes ospB. The 2 kb mRNA is post-transcriptionally processed in the intercistronic ospB-apy region, leading to the considerable accumulation of a more stable 1 kb apy-specific mRNA (half-life of 2.2+/-0.3 min, versus 27+/-4 s for the 2 kb transcript). Upon temperature induction, peak expression of the ospB-apy operon occurs when bacteria enter into the late phases of bacterial growth, where the apy-specific transcript was found to be much more prevalent if compared to the ospB-apy transcript.
- Published
- 2002
- Full Text
- View/download PDF
43. Mex67p mediates nuclear export of a variety of RNA polymerase II transcripts.
- Author
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Hurt E, Strässer K, Segref A, Bailer S, Schlaich N, Presutti C, Tollervey D, and Jansen R
- Subjects
- Adenosine Triphosphatases, Biological Transport, HSP70 Heat-Shock Proteins metabolism, Heat-Shock Proteins biosynthesis, Heat-Shock Proteins genetics, Heat-Shock Response, Membrane Proteins genetics, Membrane Proteins metabolism, Mutation, Nuclear Proteins genetics, Phosphoglycerate Kinase genetics, Phosphoglycerate Kinase metabolism, RNA Precursors metabolism, RNA-Binding Proteins genetics, Saccharomyces cerevisiae, Transcription Factors genetics, Transcription Factors metabolism, Cell Nucleus metabolism, DNA-Binding Proteins, Nuclear Pore Complex Proteins, Nuclear Proteins metabolism, Nucleocytoplasmic Transport Proteins, RNA Polymerase II metabolism, RNA, Messenger metabolism, RNA-Binding Proteins metabolism, Repressor Proteins, Saccharomyces cerevisiae Proteins
- Abstract
Mex67p is essential for nuclear poly(A)(+) RNA export in yeast, but which specific transcripts are transported by Mex67p is not known. We observed that thermosensitive mex67-5 cells do not produce a heat shock response at 37 degrees C but will induce heat shock proteins (Hsp) (e.g. Hsp104p and Hsp70p) when shifted back from the restrictive to permissive temperature (30 degrees C). This memory of a previous heat stress in mex67-5 cells could be explained if HSP mRNAs accumulated inside the nucleus during heat shock and were exported and translated in the cytoplasm on return to the permissive temperature. To test this hypothesis, nuclear export of heat shock mRNAs was directly analyzed by in situ hybridization using fluorescent-labeled oligonucleotide probes specific for SSA transcripts. This revealed that Mex67p is required for nuclear export of heat shock mRNAs. Furthermore, other polymerase II transcripts encoding the transcriptional repressor ASH1 and the glycolytic enzyme PGK1 are shown to require Mex67p for their export into the cytoplasm. Thus, Mex67p is an mRNA export factor for a broad range of polymerase II transcripts.
- Published
- 2000
- Full Text
- View/download PDF
44. Processing of the intron-encoded U18 small nucleolar RNA in the yeast Saccharomyces cerevisiae relies on both exo- and endonucleolytic activities.
- Author
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Villa T, Ceradini F, Presutti C, and Bozzoni I
- Subjects
- DNA-Binding Proteins metabolism, Exodeoxyribonuclease V, Fungal Proteins genetics, Fungal Proteins metabolism, Peptide Elongation Factor 1, Peptide Elongation Factors genetics, Peptide Elongation Factors metabolism, RNA Precursors metabolism, RNA Splicing, Ribonucleoproteins genetics, Ribonucleoproteins metabolism, Endodeoxyribonucleases metabolism, Exodeoxyribonucleases metabolism, Exoribonucleases metabolism, Introns, RNA, Small Nuclear genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins
- Abstract
Many small nucleolar RNAs (snoRNAs) are encoded within introns of protein-encoding genes and are released by processing of their host pre-mRNA. We have investigated the mechanism of processing of the yeast U18 snoRNA, which is found in the intron of the gene coding for translational elongation factor EF-1beta. We have focused our analysis on the relationship between splicing of the EF-1beta pre-mRNA and production of the mature snoRNA. Mutations inhibiting splicing of the EF-1beta pre-mRNA have been shown to produce normal U18 snoRNA levels together with the accumulation of intermediates deriving from the pre-mRNA, thus indicating that the precursor is an efficient processing substrate. Inhibition of 5'-->3' exonucleases obtained by insertion of G cassettes or by the use of a rat1-1 xrn1Delta mutant strain does not impair U18 release. In the Exo- strain, 3' cutoff products, diagnostic of an endonuclease-mediated processing pathway, were detected. Our data indicate that biosynthesis of the yeast U18 snoRNA relies on two different pathways, depending on both exonucleolytic and endonucleolytic activities: a major processing pathway based on conversion of the debranched intron and a minor one acting by endonucleolytic cleavage of the pre-mRNA.
- Published
- 1998
- Full Text
- View/download PDF
45. Self-cleaving motifs are found in close proximity to the sites utilized for U16 snoRNA processing.
- Author
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Prislei S, Fatica A, De Gregorio E, Arese M, Fragapane P, Caffarelli E, Presutti C, and Bozzoni I
- Subjects
- Animals, Base Sequence, Chromosome Mapping, DNA Transposable Elements genetics, Gene Deletion, Molecular Sequence Data, RNA, Catalytic genetics, Xenopus genetics
- Abstract
A class of small nucleolar RNAs (snoRNAs) is encoded in introns of protein-coding genes. The U16 snoRNA belongs to this class; it is encoded in the third intron of the Xenopus laevis (Xl) L1 ribosomal protein encoding gene and is released from the pre-mRNA by processing both in vivo and in vitro systems. In this paper, we show that in close proximity to the U16 snoRNA processing sites, sequences displaying self-cleaving activity are present. These elements are conserved in the two copies of the Xl L1 and in the single copy of the X. tropicalis L1. The catalytic activity corresponds to that already described for the minimal hairpin ribozyme [Dange et al., Science 242 (1990) 585-588]; it is Mn(2+)-dependent, produces 2'-3' cyclic phosphate and 5'-OH termini and comprises an essential GAAA element. Here we show that the 2'-OH group of the G residue is essential for catalysis.
- Published
- 1995
- Full Text
- View/download PDF
46. Identification of the cis-elements mediating the autogenous control of ribosomal protein L2 mRNA stability in yeast.
- Author
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Presutti C, Villa T, Hall D, Pertica C, and Bozzoni I
- Subjects
- Base Sequence, Endoribonucleases metabolism, Exoribonucleases metabolism, Feedback, Gene Expression Regulation, Fungal genetics, Introns genetics, Molecular Sequence Data, Poly G, RNA Precursors metabolism, RNA Splicing, RNA, Fungal genetics, RNA, Messenger genetics, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins metabolism, Regulatory Sequences, Nucleic Acid genetics, Sequence Deletion, beta-Galactosidase genetics, RNA Processing, Post-Transcriptional genetics, RNA, Fungal metabolism, RNA, Messenger metabolism, Ribosomal Proteins genetics, Saccharomyces cerevisiae genetics
- Abstract
The ribosomal protein L2 (rpL2) of Saccharomyces cerevisiae regulates the accumulation of its own mRNA by a feedback mechanism. An RNA sequence is responsible for this control, initially characterized as a 360 nucleotide-long region, localized at the 5' end of the transcript. This region, fused to an unrelated coding sequence, is able to down-regulate the accumulation of the chimeric transcript when increased levels of rpL2 are induced in the cell. The target regulatory region also responds to regulation when inserted inside an intron, demonstrating that the control process can take place inside the nucleus. Deletion analysis from the 5' and 3' borders have restricted the responsive region to approximately 200 nt. The insertion of a poly-G cassette downstream of the regulatory region allowed the identification of truncated 3' cut-off poly(A)+ RNA molecules. The parallel identification of cut-off molecules containing the 5' portion of the transcript allowed us to deduce that the truncated products originate by endonucleolytic cleavage. Altogether, these results are consistent with a mechanism by which the presence of excess amounts of rpL2 in the cell triggers its own mRNA to a degradative pathway; this involves an initial endonucleolytic cleavage that is followed by exonucleolytic trimming. Such a regulatory mechanism shows interesting analogies with the translational regulation of r-proteins in Escherichia coli.
- Published
- 1995
- Full Text
- View/download PDF
47. Two different snoRNAs are encoded in introns of amphibian and human L1 ribosomal protein genes.
- Author
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Prislei S, Michienzi A, Presutti C, Fragapane P, and Bozzoni I
- Subjects
- Animals, Base Sequence, Blotting, Northern, Cloning, Molecular, Conserved Sequence, DNA, Humans, Microinjections, Molecular Sequence Data, Oocytes, Phylogeny, Xenopus, Xenopus laevis, Introns, RNA genetics, Ribonucleoproteins, Small Nuclear genetics, Ribosomal Proteins genetics
- Abstract
We previously reported that the third intron of the X.laevis L1 ribosomal protein gene encodes for a snoRNA called U16. Here we show that four different introns of the same gene contain another previously uncharacterized snoRNA (U18) which is associated with fibrillarin in the nucleolus and which originates by processing of the pre-mRNA. The pathway of U18 RNA release from the pre-mRNA is the same as the one described for U16: primary endonucleolytic cleavages upstream and downstream of the U18 coding region produce a pre-U18 RNA which is subsequently trimmed to the mature form. Both the gene organization and processing of U18 are conserved in the corresponding genes of X.tropicalis and H.sapiens. The L1 gene thus has a composite structure, highly conserved in evolution, in which sequences coding for a ribosomal protein are intermingled with sequences coding for two different snoRNAs. The nucleolar localization of these different components suggests some common function on ribosome biosynthesis.
- Published
- 1993
- Full Text
- View/download PDF
48. The primary sequence of the Schizosaccharomyces pombe protein homologous to S.cerevisiae ribosomal protein L2.
- Author
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Presutti C, Villa T, and Bozzoni I
- Subjects
- Amino Acid Sequence, Fungal Proteins genetics, Molecular Sequence Data, Sequence Homology, Amino Acid, Ribosomal Proteins genetics, Saccharomyces cerevisiae genetics, Schizosaccharomyces genetics
- Published
- 1993
- Full Text
- View/download PDF
49. The mechanisms controlling ribosomal protein L1 pre-mRNA splicing are maintained in evolution and rely on conserved intron sequences.
- Author
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Prislei S, Sperandio S, Fragapane P, Caffarelli E, Presutti C, and Bozzoni I
- Subjects
- Animals, Base Sequence, Biological Evolution, Cloning, Molecular, Molecular Sequence Data, Mutagenesis genetics, Nucleic Acid Conformation, RNA Precursors genetics, RNA Splicing genetics, Ribosomal Proteins metabolism, Xenopus laevis metabolism, Introns genetics, RNA Precursors metabolism, RNA Splicing physiology, Ribosomal Proteins genetics, Xenopus laevis genetics
- Abstract
Sequences corresponding to the third intron of the X.laevis L1 ribosomal protein gene were isolated from the second copy of the X.laevis gene and from the single copy of X.tropicalis. Sequence comparison revealed that the three introns share an unusual sequence conservation which spans a region of 110 nucleotides. In addition, they have the same suboptimal 5' splice sites. The three introns show similar features upon oocyte microinjection: they have very low splicing efficiency and undergo the same site specific cleavages which lead to the accumulation of truncated molecules. Computer analysis and RNAse digestions have allowed to assign to the conserved region a specific secondary structure. Mutational analysis has shown that this structure is important for conferring the cleavage phenotype to these three introns. Competition experiments show that the cleavage phenotype can be prevented by coinjection of excess amounts of homologous sequences.
- Published
- 1992
- Full Text
- View/download PDF
50. The ribosomal protein L2 in S. cerevisiae controls the level of accumulation of its own mRNA.
- Author
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Presutti C, Ciafré SA, and Bozzoni I
- Subjects
- Blotting, Northern, Blotting, Southern, DNA, Fungal genetics, Gene Expression Regulation, Fungal, Plasmids, RNA Processing, Post-Transcriptional, RNA, Fungal genetics, RNA, Messenger genetics, Ribosomal Proteins genetics, Transcription, Genetic, RNA, Fungal metabolism, RNA, Messenger metabolism, Ribosomal Proteins metabolism, Saccharomyces cerevisiae metabolism
- Abstract
The expression of the yeast L2 r-protein gene is controlled at the level of mRNA accumulation. The product of the gene appears to participate in this regulation by an autogenous feedback mechanism. This control does not operate at the level of transcription but instead affects L2 mRNA accumulation. This autogenous regulation of mRNA accumulation provides an interesting analogy to the autogenous translational regulation of r-proteins in Escherichia coli.
- Published
- 1991
- Full Text
- View/download PDF
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