22 results on '"Ajore R"'
Search Results
2. Immobilizing antibodies on piezoelectric quartz crystal for diagnostic applications.
- Author
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Vashist, S.K., Tewari, R., Ajore, R., Bajpai, R.P., and Bharadwaj, L.M.
- Published
- 2005
- Full Text
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3. Immobilizing antibodies on piezoelectric quartz crystal for diagnostic applications
- Author
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Vashist, S.K., primary, Tewari, R., additional, Ajore, R., additional, Bajpai, R.P., additional, and Bharadwaj, L.M., additional
- Full Text
- View/download PDF
4. Deciphering the genetics and mechanisms of predisposition to multiple myeloma.
- Author
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Went M, Duran-Lozano L, Halldorsson GH, Gunnell A, Ugidos-Damboriena N, Law P, Ekdahl L, Sud A, Thorleifsson G, Thodberg M, Olafsdottir T, Lamarca-Arrizabalaga A, Cafaro C, Niroula A, Ajore R, Lopez de Lapuente Portilla A, Ali Z, Pertesi M, Goldschmidt H, Stefansdottir L, Kristinsson SY, Stacey SN, Love TJ, Rognvaldsson S, Hajek R, Vodicka P, Pettersson-Kymmer U, Späth F, Schinke C, Van Rhee F, Sulem P, Ferkingstad E, Hjorleifsson Eldjarn G, Mellqvist UH, Jonsdottir I, Morgan G, Sonneveld P, Waage A, Weinhold N, Thomsen H, Försti A, Hansson M, Juul-Vangsted A, Thorsteinsdottir U, Hemminki K, Kaiser M, Rafnar T, Stefansson K, Houlston R, and Nilsson B
- Subjects
- Humans, Mendelian Randomization Analysis, B-Lymphocytes immunology, B-Lymphocytes metabolism, Case-Control Studies, Transmembrane Activator and CAML Interactor Protein genetics, Male, Telomere genetics, Multiple Myeloma genetics, Genetic Predisposition to Disease, Genome-Wide Association Study, B-Cell Maturation Antigen genetics, Polymorphism, Single Nucleotide
- Abstract
Multiple myeloma (MM) is an incurable malignancy of plasma cells. Epidemiological studies indicate a substantial heritable component, but the underlying mechanisms remain unclear. Here, in a genome-wide association study totaling 10,906 cases and 366,221 controls, we identify 35 MM risk loci, 12 of which are novel. Through functional fine-mapping and Mendelian randomization, we uncover two causal mechanisms for inherited MM risk: longer telomeres; and elevated levels of B-cell maturation antigen (BCMA) and interleukin-5 receptor alpha (IL5RA) in plasma. The largest increase in BCMA and IL5RA levels is mediated by the risk variant rs34562254-A at TNFRSF13B. While individuals with loss-of-function variants in TNFRSF13B develop B-cell immunodeficiency, rs34562254-A exerts a gain-of-function effect, increasing MM risk through amplified B-cell responses. Our results represent an analysis of genetic MM predisposition, highlighting causal mechanisms contributing to MM development., (© 2024. The Author(s).)
- Published
- 2024
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5. Genome-wide CRISPR/Cas9 screen identifies regulators of BCMA expression on multiple myeloma cells.
- Author
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Ajore R, Mattsson J, Pertesi M, Ekdahl L, Ali Z, Hansson M, and Nilsson B
- Subjects
- Humans, B-Cell Maturation Antigen, CRISPR-Cas Systems, Cell Line, Tumor, Multiple Myeloma genetics, Multiple Myeloma metabolism
- Published
- 2024
- Full Text
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6. Author Correction: Functional dissection of inherited non-coding variation influencing multiple myeloma risk.
- Author
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Ajore R, Niroula A, Pertesi M, Cafaro C, Thodberg M, Went M, Bao EL, Duran-Lozano L, Lopez de Lapuente Portilla A, Olafsdottir T, Ugidos-Damboriena N, Magnusson O, Samur M, Lareau CA, Halldorsson GH, Thorleifsson G, Norddahl GL, Gunnarsdottir K, Försti A, Goldschmidt H, Hemminki K, van Rhee F, Kimber S, Sperling AS, Kaiser M, Anderson K, Jonsdottir I, Munshi N, Rafnar T, Waage A, Weinhold N, Thorsteinsdottir U, Sankaran VG, Stefansson K, Houlston R, and Nilsson B
- Published
- 2022
- Full Text
- View/download PDF
7. Functional dissection of inherited non-coding variation influencing multiple myeloma risk.
- Author
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Ajore R, Niroula A, Pertesi M, Cafaro C, Thodberg M, Went M, Bao EL, Duran-Lozano L, Lopez de Lapuente Portilla A, Olafsdottir T, Ugidos-Damboriena N, Magnusson O, Samur M, Lareau CA, Halldorsson GH, Thorleifsson G, Norddahl GL, Gunnarsdottir K, Försti A, Goldschmidt H, Hemminki K, van Rhee F, Kimber S, Sperling AS, Kaiser M, Anderson K, Jonsdottir I, Munshi N, Rafnar T, Waage A, Weinhold N, Thorsteinsdottir U, Sankaran VG, Stefansson K, Houlston R, and Nilsson B
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Adaptor Proteins, Signal Transducing immunology, Antineoplastic Combined Chemotherapy Protocols, B-Lymphocytes immunology, Base Sequence, Cell Cycle Proteins genetics, Cell Cycle Proteins immunology, Chromatin chemistry, Chromatin immunology, Chromosomal Proteins, Non-Histone genetics, Chromosomal Proteins, Non-Histone immunology, DNA, Intergenic immunology, Gene Expression Regulation, Neoplastic, Guanine Nucleotide Exchange Factors genetics, Guanine Nucleotide Exchange Factors immunology, Humans, Inheritance Patterns, Multiple Myeloma drug therapy, Multiple Myeloma immunology, Multiple Myeloma pathology, Neoplasm Proteins immunology, Plasma Cells immunology, Polymorphism, Genetic, Primary Cell Culture, Quantitative Trait Loci, Repressor Proteins genetics, Repressor Proteins immunology, Risk Assessment, Transcriptional Elongation Factors genetics, Transcriptional Elongation Factors immunology, B-Lymphocytes pathology, DNA, Intergenic genetics, Genetic Predisposition to Disease, Multiple Myeloma genetics, Neoplasm Proteins genetics, Plasma Cells pathology
- Abstract
Thousands of non-coding variants have been associated with increased risk of human diseases, yet the causal variants and their mechanisms-of-action remain obscure. In an integrative study combining massively parallel reporter assays (MPRA), expression analyses (eQTL, meQTL, PCHiC) and chromatin accessibility analyses in primary cells (caQTL), we investigate 1,039 variants associated with multiple myeloma (MM). We demonstrate that MM susceptibility is mediated by gene-regulatory changes in plasma cells and B-cells, and identify putative causal variants at six risk loci (SMARCD3, WAC, ELL2, CDCA7L, CEP120, and PREX1). Notably, three of these variants co-localize with significant plasma cell caQTLs, signaling the presence of causal activity at these precise genomic positions in an endogenous chromosomal context in vivo. Our results provide a systematic functional dissection of risk loci for a hematologic malignancy., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
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8. Author Correction: Germline variants at SOHLH2 influence multiple myeloma risk.
- Author
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Duran-Lozano L, Thorleifsson G, Lopez de Lapuente Portilla A, Niroula A, Went M, Thodberg M, Pertesi M, Ajore R, Cafaro C, Olason PI, Stefansdottir L, Bragi Walters G, Halldorsson GH, Turesson I, Kaiser MF, Weinhold N, Abildgaard N, Andersen NF, Mellqvist UH, Waage A, Juul-Vangsted A, Thorsteinsdottir U, Hansson M, Houlston R, Rafnar T, Stefansson K, and Nilsson B
- Published
- 2021
- Full Text
- View/download PDF
9. Germline variants at SOHLH2 influence multiple myeloma risk.
- Author
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Duran-Lozano L, Thorleifsson G, Lopez de Lapuente Portilla A, Niroula A, Went M, Thodberg M, Pertesi M, Ajore R, Cafaro C, Olason PI, Stefansdottir L, Bragi Walters G, Halldorsson GH, Turesson I, Kaiser MF, Weinhold N, Abildgaard N, Andersen NF, Mellqvist UH, Waage A, Juul-Vangsted A, Thorsteinsdottir U, Hansson M, Houlston R, Rafnar T, Stefansson K, and Nilsson B
- Subjects
- Aged, Female, Gene Frequency, Genetic Predisposition to Disease, Genome-Wide Association Study, Germ Cells metabolism, Germ-Line Mutation, Humans, Linkage Disequilibrium, Male, Polymorphism, Single Nucleotide, Basic Helix-Loop-Helix Transcription Factors genetics, Multiple Myeloma genetics
- Abstract
Multiple myeloma (MM) is caused by the uncontrolled, clonal expansion of plasma cells. While there is epidemiological evidence for inherited susceptibility, the molecular basis remains incompletely understood. We report a genome-wide association study totalling 5,320 cases and 422,289 controls from four Nordic populations, and find a novel MM risk variant at SOHLH2 at 13q13.3 (risk allele frequency = 3.5%; odds ratio = 1.38; P = 2.2 × 10
-14 ). This gene encodes a transcription factor involved in gametogenesis that is normally only weakly expressed in plasma cells. The association is represented by 14 variants in linkage disequilibrium. Among these, rs75712673 maps to a genomic region with open chromatin in plasma cells, and upregulates SOHLH2 in this cell type. Moreover, rs75712673 influences transcriptional activity in luciferase assays, and shows a chromatin looping interaction with the SOHLH2 promoter. Our work provides novel insight into MM susceptibility.- Published
- 2021
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10. Accelerating target deconvolution for therapeutic antibody candidates using highly parallelized genome editing.
- Author
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Mattsson J, Ekdahl L, Junghus F, Ajore R, Erlandsson E, Niroula A, Pertesi M, Frendéus B, Teige I, and Nilsson B
- Subjects
- Antibodies metabolism, CRISPR-Cas Systems genetics, Cell Line, Tumor, Cell Survival genetics, Cell Survival physiology, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Humans, Gene Editing
- Abstract
Therapeutic antibodies are transforming the treatment of cancer and autoimmune diseases. Today, a key challenge is finding antibodies against new targets. Phenotypic discovery promises to achieve this by enabling discovery of antibodies with therapeutic potential without specifying the molecular target a priori. Yet, deconvoluting the targets of phenotypically discovered antibodies remains a bottleneck; efficient deconvolution methods are needed for phenotypic discovery to reach its full potential. Here, we report a comprehensive investigation of a target deconvolution approach based on pooled CRISPR/Cas9. Applying this approach within three real-world phenotypic discovery programs, we rapidly deconvolute the targets of 38 of 39 test antibodies (97%), a success rate far higher than with existing approaches. Moreover, the approach scales well, requires much less work, and robustly identifies antibodies against the major histocompatibility complex. Our data establish CRISPR/Cas9 as a highly efficient target deconvolution approach, with immediate implications for the development of antibody-based drugs.
- Published
- 2021
- Full Text
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11. MPRAscore: robust and non-parametric analysis of massively parallel reporter assays.
- Author
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Niroula A, Ajore R, and Nilsson B
- Subjects
- Alleles, Biological Assay, Software
- Abstract
Motivation: Massively parallel reporter assays (MPRA) enable systematic screening of DNA sequence variants for effects on transcriptional activity. However, convenient analysis tools are still needed., Results: We introduce MPRAscore, a novel tool to infer allele-specific effects on transcription from MPRA data. MPRAscore uses a weighted, variance-regularized method to calculate variant effect sizes robustly, and a permutation approach to test for significance without assuming normality or independence., Availability and Implementation: Source code (C++), precompiled binaries and data used in the paper at https://github.com/abhisheknrl/MPRAscore and https://www.ncbi.nlm.nih.gov/bioproject/PRJNA554195., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2019
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12. The multiple myeloma risk allele at 5q15 lowers ELL2 expression and increases ribosomal gene expression.
- Author
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Ali M, Ajore R, Wihlborg AK, Niroula A, Swaminathan B, Johnsson E, Stephens OW, Morgan G, Meissner T, Turesson I, Goldschmidt H, Mellqvist UH, Gullberg U, Hansson M, Hemminki K, Nahi H, Waage A, Weinhold N, and Nilsson B
- Subjects
- Bone Marrow pathology, Cell Line, Tumor, Datasets as Topic, Down-Regulation, Gene Expression Profiling, Gene Knockout Techniques, Genetic Predisposition to Disease, Humans, Multiple Myeloma pathology, Plasma Cells metabolism, Polymorphism, Single Nucleotide, Quantitative Trait Loci genetics, Ribosomal Proteins metabolism, Ribosomes genetics, Ribosomes metabolism, Transcriptional Elongation Factors metabolism, Up-Regulation, Alleles, Chromosomes, Human, Pair 5 genetics, Gene Expression Regulation, Neoplastic, Multiple Myeloma genetics, Ribosomal Proteins genetics, Transcriptional Elongation Factors genetics
- Abstract
Recently, we identified ELL2 as a susceptibility gene for multiple myeloma (MM). To understand its mechanism of action, we performed expression quantitative trait locus analysis in CD138
+ plasma cells from 1630 MM patients from four populations. We show that the MM risk allele lowers ELL2 expression in these cells (Pcombined = 2.5 × 10-27 ; βcombined = -0.24 SD), but not in peripheral blood or other tissues. Consistent with this, several variants representing the MM risk allele map to regulatory genomic regions, and three yield reduced transcriptional activity in plasmocytoma cell lines. One of these (rs3777189-C) co-locates with the best-supported lead variants for ELL2 expression and MM risk, and reduces binding of MAFF/G/K family transcription factors. Moreover, further analysis reveals that the MM risk allele associates with upregulation of gene sets related to ribosome biogenesis, and knockout/knockdown and rescue experiments in plasmocytoma cell lines support a cause-effect relationship. Our results provide mechanistic insight into MM predisposition.- Published
- 2018
- Full Text
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13. Identification of sequence variants influencing immunoglobulin levels.
- Author
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Jonsson S, Sveinbjornsson G, de Lapuente Portilla AL, Swaminathan B, Plomp R, Dekkers G, Ajore R, Ali M, Bentlage AEH, Elmér E, Eyjolfsson GI, Gudjonsson SA, Gullberg U, Gylfason A, Halldorsson BV, Hansson M, Holm H, Johansson Å, Johnsson E, Jonasdottir A, Ludviksson BR, Oddsson A, Olafsson I, Olafsson S, Sigurdardottir O, Sigurdsson A, Stefansdottir L, Masson G, Sulem P, Wuhrer M, Wihlborg AK, Thorleifsson G, Gudbjartsson DF, Thorsteinsdottir U, Vidarsson G, Jonsdottir I, Nilsson B, and Stefansson K
- Subjects
- Cohort Studies, Female, Genetic Predisposition to Disease, Genome-Wide Association Study, Hematopoiesis genetics, Humans, Iceland, Immunity, Humoral genetics, Immunoglobulin Class Switching genetics, Immunoglobulin Isotypes genetics, Male, Polymorphism, Single Nucleotide, Sweden, Genetic Variation, Immunoglobulins genetics
- Abstract
Immunoglobulins are the effector molecules of the adaptive humoral immune system. In a genome-wide association study of 19,219 individuals, we found 38 new variants and replicated 5 known variants associating with IgA, IgG or IgM levels or with composite immunoglobulin traits, accounted for by 32 loci. Variants at these loci also affect the risk of autoimmune diseases and blood malignancies and influence blood cell development. Notable associations include a rare variant at RUNX3 decreasing IgA levels by shifting isoform proportions (rs188468174[C>T]: P = 8.3 × 10
-55 , β = -0.90 s.d.), a rare in-frame deletion in FCGR2B abolishing IgG binding to the encoded receptor (p.Asn106del: P = 4.2 × 10-8 , β = 1.03 s.d.), four IGH locus variants influencing class switching, and ten new associations with the HLA region. Our results provide new insight into the regulation of humoral immunity.- Published
- 2017
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14. Deletion of ribosomal protein genes is a common vulnerability in human cancer, especially in concert with TP53 mutations.
- Author
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Ajore R, Raiser D, McConkey M, Jöud M, Boidol B, Mar B, Saksena G, Weinstock DM, Armstrong S, Ellis SR, Ebert BL, and Nilsson B
- Subjects
- Cell Line, Tumor, Gene Expression Regulation, Neoplastic, Humans, Gene Deletion, Mutation, Neoplasms pathology, Ribosomal Proteins genetics, Tumor Suppressor Protein p53 genetics
- Abstract
Heterozygous inactivating mutations in ribosomal protein genes (RPGs) are associated with hematopoietic and developmental abnormalities, activation of p53, and altered risk of cancer in humans and model organisms. Here we performed a large-scale analysis of cancer genome data to examine the frequency and selective pressure of RPG lesions across human cancers. We found that hemizygous RPG deletions are common, occurring in about 43% of 10,744 cancer specimens and cell lines. Consistent with p53-dependent negative selection, such lesions are underrepresented in TP53 -intact tumors ( P ≪ 10
-10 ), and shRNA-mediated knockdown of RPGs activated p53 in TP53 -wild-type cells. In contrast, we did not see negative selection of RPG deletions in TP53 -mutant tumors. RPGs are conserved with respect to homozygous deletions, and shRNA screening data from 174 cell lines demonstrate that further suppression of hemizygously deleted RPGs inhibits cell growth. Our results establish RPG haploinsufficiency as a strikingly common vulnerability of human cancers that associates with TP53 mutations and could be targetable therapeutically., (© 2017 The Authors. Published under the terms of the CC BY 4.0 license.)- Published
- 2017
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15. SMIM1 variants rs1175550 and rs143702418 independently modulate Vel blood group antigen expression.
- Author
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Christophersen MK, Jöud M, Ajore R, Vege S, Ljungdahl KW, Westhoff CM, Olsson ML, Storry JR, and Nilsson B
- Subjects
- Alleles, Base Sequence, Gene Frequency genetics, Genotype, Humans, Introns genetics, Luciferases metabolism, Membrane Proteins metabolism, Protein Binding, T-Cell Acute Lymphocytic Leukemia Protein 1 metabolism, Transcription, Genetic, Blood Group Antigens metabolism, Membrane Proteins genetics, Polymorphism, Single Nucleotide genetics
- Abstract
The Vel blood group antigen is expressed on the red blood cells of most individuals. Recently, we described that homozygosity for inactivating mutations in SMIM1 defines the rare Vel-negative phenotype. Still, Vel-positive individuals show great variability in Vel antigen expression, creating a risk for Vel blood typing errors and transfusion reactions. We fine-mapped the regulatory region located in SMIM1 intron 2 in Swedish blood donors, and observed a strong correlation between expression and rs1175550 as well as with a previously unreported tri-nucleotide insertion (rs143702418; C > CGCA). While the two variants are tightly linked in Caucasians, we separated their effects in African Americans, and found that rs1175550G and to a lesser extent rs143702418C independently increase SMIM1 and Vel antigen expression. Gel shift and luciferase assays indicate that both variants are transcriptionally active, and we identified binding of the transcription factor TAL1 as a potential mediator of the increased expression associated with rs1175550G. Our results provide insight into the regulatory logic of Vel antigen expression, and extend the set of markers for genetic Vel blood group typing.
- Published
- 2017
- Full Text
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16. Variants in ELL2 influencing immunoglobulin levels associate with multiple myeloma.
- Author
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Swaminathan B, Thorleifsson G, Jöud M, Ali M, Johnsson E, Ajore R, Sulem P, Halvarsson BM, Eyjolfsson G, Haraldsdottir V, Hultman C, Ingelsson E, Kristinsson SY, Kähler AK, Lenhoff S, Masson G, Mellqvist UH, Månsson R, Nelander S, Olafsson I, Sigurðardottir O, Steingrimsdóttir H, Vangsted A, Vogel U, Waage A, Nahi H, Gudbjartsson DF, Rafnar T, Turesson I, Gullberg U, Stefánsson K, Hansson M, Thorsteinsdóttir U, and Nilsson B
- Subjects
- Genetic Predisposition to Disease, Genome-Wide Association Study, Humans, Intracellular Signaling Peptides and Proteins, Meningitis, Bacterial genetics, Immunoglobulin A blood, Immunoglobulin G blood, Multiple Myeloma genetics, Proteins genetics, Transcriptional Elongation Factors genetics
- Abstract
Multiple myeloma (MM) is characterized by an uninhibited, clonal growth of plasma cells. While first-degree relatives of patients with MM show an increased risk of MM, the genetic basis of inherited MM susceptibility is incompletely understood. Here we report a genome-wide association study in the Nordic region identifying a novel MM risk locus at ELL2 (rs56219066T; odds ratio (OR)=1.25; P=9.6 × 10(-10)). This gene encodes a stoichiometrically limiting component of the super-elongation complex that drives secretory-specific immunoglobulin mRNA production and transcriptional regulation in plasma cells. We find that the MM risk allele harbours a Thr298Ala missense variant in an ELL2 domain required for transcription elongation. Consistent with a hypomorphic effect, we find that the MM risk allele also associates with reduced levels of immunoglobulin A (IgA) and G (IgG) in healthy subjects (P=8.6 × 10(-9) and P=6.4 × 10(-3), respectively) and, potentially, with an increased risk of bacterial meningitis (OR=1.30; P=0.0024).
- Published
- 2015
- Full Text
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17. Myeloid translocation gene-16 co-repressor promotes degradation of hypoxia-inducible factor 1.
- Author
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Kumar P, Gullberg U, Olsson I, and Ajore R
- Subjects
- Aryl Hydrocarbon Receptor Nuclear Translocator genetics, Aryl Hydrocarbon Receptor Nuclear Translocator metabolism, B-Lymphocytes cytology, Cell Line, Tumor, Chromatin Immunoprecipitation, Gene Expression Regulation, Humans, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Hypoxia-Inducible Factor-Proline Dioxygenases genetics, Hypoxia-Inducible Factor-Proline Dioxygenases metabolism, Oxidative Phosphorylation, Phosphofructokinase-2 genetics, Phosphofructokinase-2 metabolism, Promoter Regions, Genetic, Proteasome Endopeptidase Complex metabolism, Protein Binding, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases metabolism, Protein Stability, Proteolysis, Pyruvate Dehydrogenase Acetyl-Transferring Kinase, Repressor Proteins metabolism, Response Elements, Signal Transduction, Tumor Suppressor Proteins metabolism, Ubiquitination, B-Lymphocytes metabolism, Glycolysis genetics, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Repressor Proteins genetics, Tumor Suppressor Proteins genetics
- Abstract
The myeloid translocation gene 16 (MTG16) co-repressor down regulates expression of multiple glycolytic genes, which are targets of the hypoxia-inducible factor 1 (HIF1) heterodimer transcription factor that is composed of oxygen-regulated labile HIF1α and stable HIF1β subunits. For this reason, we investigated whether MTG16 might regulate HIF1 negatively contributing to inhibition of glycolysis and stimulation of mitochondrial respiration. A doxycycline Tet-On system was used to control levels of MTG16 in B-lymphoblastic Raji cells. Results from co-association studies revealed MTG16 to interact with HIF1α. The co-association required intact N-terminal MTG16 residues including Nervy Homology Region 1 (NHR1). Furthermore, electrophoretic mobility shift assays demonstrated an association of MTG16 with hypoxia response elements (HREs) in PFKFB3, PFKFB4 and PDK1 promoters in-vitro. Results from chromatin immunoprecipitation assays revealed co-occupancy of these and other glycolytic gene promoters by HIF1α, HIF1β and MTG16 in agreement with possible involvement of these proteins in regulation of glycolytic target genes. In addition, MTG16 interacted with prolyl hydroxylase D2 and promoted ubiquitination and proteasomal degradation of HIF1α. Our findings broaden the area of MTG co-repressor functions and reveal MTG16 to be part of a protein complex that controls the levels of HIF1α.
- Published
- 2015
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18. The transcriptional co-repressor myeloid translocation gene 16 inhibits glycolysis and stimulates mitochondrial respiration.
- Author
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Kumar P, Sharoyko VV, Spégel P, Gullberg U, Mulder H, Olsson I, and Ajore R
- Subjects
- Cell Hypoxia, Cell Line, Tumor, Cell Proliferation, Gene Expression Regulation, Humans, MAP Kinase Signaling System, Mitogen-Activated Protein Kinase 8 metabolism, Oxygen Consumption, Phosphofructokinase-2 genetics, Phosphofructokinase-2 metabolism, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases metabolism, Pyruvate Dehydrogenase Acetyl-Transferring Kinase, Glycolysis, Mitochondria metabolism, Repressor Proteins genetics, Repressor Proteins metabolism, Tumor Suppressor Proteins genetics, Tumor Suppressor Proteins metabolism
- Abstract
The myeloid translocation gene 16 product MTG16 is found in multiple transcription factor-containing complexes as a regulator of gene expression implicated in development and tumorigenesis. A stable Tet-On system for doxycycline-dependent expression of MTG16 was established in B-lymphoblastoid Raji cells to unravel its molecular functions in transformed cells. A noticeable finding was that expression of certain genes involved in tumor cell metabolism including 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 and 4 (PFKFB3 and PFKFB4), and pyruvate dehydrogenase kinase isoenzyme 1 (PDK1) was rapidly diminished when MTG16 was expressed. Furthermore, hypoxia-stimulated production of PFKFB3, PFKFB4 and PDK1 was inhibited by MTG16 expression. The genes in question encode key regulators of glycolysis and its coupling to mitochondrial metabolism and are commonly found to be overexpressed in transformed cells. The MTG16 Nervy Homology Region 2 (NHR2) oligomerization domain and the NHR3 protein-protein interaction domain were required intact for inhibition of PFKFB3, PFKFB4 and PDK1 expression to occur. Expression of MTG16 reduced glycolytic metabolism while mitochondrial respiration and formation of reactive oxygen species increased. The metabolic changes were paralleled by increased phosphorylation of mitogen-activated protein kinases, reduced levels of amino acids and inhibition of proliferation with a decreased fraction of cells in S-phase. Overall, our findings show that MTG16 can serve as a brake on glycolysis, a stimulator of mitochondrial respiration and an inhibitor of cell proliferation. Hence, elevation of MTG16 might have anti-tumor effect.
- Published
- 2013
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19. The leukemia associated nuclear corepressor ETO homologue genes MTG16 and MTGR1 are regulated differently in hematopoietic cells.
- Author
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Ajore R, Kumar P, Dhanda RS, Gullberg U, and Olsson I
- Subjects
- Animals, Base Sequence, Basic Helix-Loop-Helix Transcription Factors metabolism, Binding Sites, Callithrix, Cell Line, Tumor, Core Binding Factor Alpha 2 Subunit metabolism, GATA Transcription Factors antagonists & inhibitors, GATA Transcription Factors genetics, GATA Transcription Factors metabolism, Gorilla gorilla, Humans, Leukemia metabolism, Leukemia pathology, Macaca mulatta, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Oncogene Proteins, Fusion metabolism, Pan troglodytes, Promoter Regions, Genetic, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, RNA Interference, RNA, Small Interfering metabolism, RUNX1 Translocation Partner 1 Protein, Rats, Repressor Proteins antagonists & inhibitors, Repressor Proteins genetics, Sp1 Transcription Factor metabolism, TATA Box genetics, Transcription Factors genetics, Transcription Factors metabolism, Tumor Suppressor Proteins antagonists & inhibitors, Tumor Suppressor Proteins genetics, U937 Cells, Gene Expression Regulation, Repressor Proteins metabolism, Tumor Suppressor Proteins metabolism
- Abstract
Background: MTG16, MTGR1 and ETO are nuclear transcriptional corepressors of the human ETO protein family. MTG16 is implicated in hematopoietic development and in controlling erythropoiesis/megakaryopoiesis. Furthermore, ETO homologue genes are 3'participants in leukemia fusions generated by chromosomal translocations responsible of hematopoietic dysregulation. We tried to identify structural and functional promoter elements of MTG16 and MTGR1 genes in order to find associations between their regulation and hematopoiesis., Results: 5' deletion examinations and luciferase reporter gene studies indicated that a 492 bp sequence upstream of the transcription start site is essential for transcriptional activity by the MTG16 promoter. The TATA- and CCAAT-less promoter with a GC box close to the start site showed strong reporter activity when examined in erythroid/megakaryocytic cells. Mutation of an evolutionary conserved GATA -301 consensus binding site repressed promoter function. Furthermore, results from in vitro antibody-enhanced electrophoretic mobility shift assay and in vivo chromatin immunoprecipitation indicated binding of GATA-1 to the GATA -301 site. A role of GATA-1 was also supported by transfection of small interfering RNA, which diminished MTG16 expression. Furthermore, expression of the transcription factor HERP2, which represses GATA-1, produced strong inhibition of the MTG16 promoter reporter consistent with a role of GATA-1 in transcriptional activation. The TATA-less and CCAAT-less MTGR1 promoter retained most of the transcriptional activity within a -308 to -207 bp region with a GC-box-rich sequence containing multiple SP1 binding sites reminiscent of a housekeeping gene with constitutive expression. However, mutations of individual SP1 binding sites did not repress promoter function; multiple active SP1 binding sites may be required to safeguard constitutive MTGR1 transcriptional activity. The observed repression of MTG16/MTGR1 promoters by the leukemia associated AML1-ETO fusion gene may have a role in hematopoietic dysfunction of leukemia., Conclusions: An evolutionary conserved GATA binding site is critical in transcriptional regulation of the MTG16 promoter. In contrast, the MTGR1 gene depends on a GC-box-rich sequence for transcriptional regulation and possible ubiquitous expression. Our results demonstrate that the ETO homologue promoters are regulated differently consistent with hematopoietic cell-type- specific expression and function.
- Published
- 2012
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20. The leukemia associated ETO nuclear repressor gene is regulated by the GATA-1 transcription factor in erythroid/megakaryocytic cells.
- Author
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Ajore R, Dhanda RS, Gullberg U, and Olsson I
- Subjects
- Animals, Base Sequence, COS Cells, Cell Line, Tumor, Chlorocebus aethiops, Core Binding Factor Alpha 2 Subunit metabolism, HL-60 Cells, Humans, Leukemia, Erythroblastic, Acute genetics, Leukemia, Erythroblastic, Acute metabolism, Leukemia, Myeloid, Acute genetics, Molecular Sequence Data, Oncogene Proteins, Fusion metabolism, Promoter Regions, Genetic, Proto-Oncogene Proteins metabolism, RUNX1 Translocation Partner 1 Protein, Transcription Factors metabolism, GATA1 Transcription Factor metabolism, Gene Expression Regulation, Megakaryocytes metabolism, Proto-Oncogene Proteins genetics, Transcription Factors genetics
- Abstract
Background: The Eight-Twenty-One (ETO) nuclear co-repressor gene belongs to the ETO homologue family also containing Myeloid Translocation Gene on chromosome 16 (MTG16) and myeloid translocation Gene-Related protein 1 (MTGR1). By chromosomal translocations ETO and MTG16 become parts of fusion proteins characteristic of morphological variants of acute myeloid leukemia. Normal functions of ETO homologues have as yet not been examined. The goal of this work was to identify structural and functional promoter elements upstream of the coding sequence of the ETO gene in order to explore lineage-specific hematopoietic expression and get hints to function., Results: A putative proximal ETO promoter was identified within 411 bp upstream of the transcription start site. Strong ETO promoter activity was specifically observed upon transfection of a promoter reporter construct into erythroid/megakaryocytic cells, which have endogeneous ETO gene activity. An evolutionary conserved region of 228 bp revealed potential cis-elements involved in transcription of ETO. Disruption of the evolutionary conserved GATA -636 consensus binding site repressed transactivation and disruption of the ETS1 -705 consensus binding site enhanced activity of the ETO promoter. The promoter was stimulated by overexpression of GATA-1 into erythroid/megakaryocytic cells. Electrophoretic mobility shift assay with erythroid/megakaryocytic cells showed specific binding of GATA-1 to the GATA -636 site. Furthermore, results from chromatin immunoprecipitation showed GATA-1 binding in vivo to the conserved region of the ETO promoter containing the -636 site. The results suggest that the GATA -636 site may have a role in activation of the ETO gene activity in cells with erythroid/megakaryocytic potential. Leukemia associated AML1-ETO strongly suppressed an ETO promoter reporter in erythroid/megakaryocytic cells., Conclusions: We demonstrate that the GATA-1 transcription factor binds and transactivates the ETO proximal promoter in an erythroid/megakaryocytic-specific manner. Thus, trans-acting factors that are essential in erythroid/megakaryocytic differentiation govern ETO expression.
- Published
- 2010
- Full Text
- View/download PDF
21. Covalent attachment of actin filaments to Tween 80 coated polystyrene beads for cargo transportation.
- Author
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Kaur H, Das T, Kumar R, Ajore R, and Bharadwaj LM
- Subjects
- Actins chemistry, Polysorbates chemistry, Polystyrenes chemistry
- Abstract
In this manuscript, a new strategy has been reported for circumscribed covalent attachment of barbed and pointed ends of actin filaments to polystyrene beads. A comparative study of attachment of actin filaments to polystyrene beads was performed by blocking functionally active sites on polystyrene beads with nonionic detergents such as Tween 20, Tween 80 and polyethylene glycol (PEG). Effective blocking of active sites was obtained with Tween 80 at 0.1% concentration. Attachment of single bundle of actin filament to bead was assessed by rotational motion of bead tailed actin in front and lateral view. Velocity of actin filaments attached to different size of beads in in-vitro motility assay was calculated to ascertain their attachments. Velocity of actin attached to 1.0 and 3.0 microm polystyrene beads was reduced to 3.0-4.0 and 0.0-1.0 microm/s, respectively as compared to free actin velocity of 4.0-6.0 microm/s. Single point attachment of actin filaments to different size of beads was assessed by decrease in sliding velocity. Present study provides insight into the actin-myosin based molecular motor systems for drug delivery and biosensors applications.
- Published
- 2008
- Full Text
- View/download PDF
22. DNA immobilization chemical interference due to aggregates study by Dip and Drop approach.
- Author
-
Ajore R, Kumar R, Kaur I, Sobti RC, and Bharadwaj LM
- Subjects
- Adsorption, Bacteriophage lambda chemistry, Base Sequence, Biocompatible Materials chemistry, DNA, Viral chemistry, Electrochemistry, Gold, Macromolecular Substances chemistry, Microelectrodes, Oligodeoxyribonucleotides chemistry, Surface Properties, Thionucleotides chemistry, DNA chemistry
- Abstract
In the present manuscript, we report the studies and observations for chemical interference due to aggregates formation during covalent immobilization of thiolated lambda-DNA between gold microelectrodes. Dip and Drop approaches were employed to study DNA immobilization using thiolated oligos (oligoA 5' GGGCGGCGACCT 3' and oligoB 5' AGGTCGCCGCCC 3'). As a result of aggregation, less interference was observed in Dip approach as compared to Drop approach. Atomic Force Microscopy (AFM) analysis of piranha treated gold surface revealed 47.5% increase in height roughness, contributing in interference by creating active sites. Cyclic voltammetry (CV) studies ascertain the multitude of adsorption states existing in long strand of DNA on surface. Surface coverage was found to be approximately 72% (1.35x10(10) molecules/cm(2)), and approximately 42% (7.89x10(9) molecules/ cm(2)) in Dip and Drop approach, respectively. Dip approach can be used as a measure to minimize interference due to aggregation.
- Published
- 2007
- Full Text
- View/download PDF
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