| Oligonucleotide decoy mimicking alphaA-crystallin-binding protein 1 binding site on mouse Col2a1 enhancer stimulates transcription from the adjacent Col2a1 promoter in chondrogenic ATDC5 cell. | |
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MedLine Citation:
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PMID: 15456958 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A 48-bp sequence element in intron 1 of the alpha1(II) collagen gene (Col2a1) acts as an enhancer of Col2a1 transcription and contains binding sites for the transcription activator SOX9 and repressor alphaA-crystallin-binding protein 1 (CRYBP1). We hypothesized that abrogating CRYBP1 binding should increase transcription from a promoter associated with the Col2a1 enhancer. We tested this hypothesis by cotransfecting an oligonucleotide (ODN) decoy for CRYBP1 and a luciferase-based reporter vector under the transcriptional control of the Col2a1 promoter linked to the 100-bp enhancer in chondrogenic ATDC5 cells. As a control, we used decoy ODN corresponding to the SOX9 binding site. Transfection with CRYBP1 decoy increased luciferase activity by >2.5-fold in the absence or presence of insulin, whereas SOX9 decoy ODN decreased luciferase activity to about 50% under similar conditions. In addition, the repressive effect of interleukin-1 on Col2a1 transcription through decreasing SOX9 messenger ribonucleic acid (mRNA) expression and increasing CRYBP1 mRNA expression, was counteracted by CRYBP1 decoy ODN. These results provide a rationale for gene therapy in degenerative joint diseases by elevating Col2a1 expression in chondrocytes through oligomimetics of repressor binding sites. |
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Authors:
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Hiroshi Yamagiwa; Yoshihiko Yamada; Mark E Bolander; Gobinda Sarkar |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Molecular biotechnology Volume: 28 ISSN: 1073-6085 ISO Abbreviation: Mol. Biotechnol. Publication Date: 2004 Sep |
Date Detail:
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Created Date: 2004-09-30 Completed Date: 2005-04-28 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 9423533 Medline TA: Mol Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 1-8 Citation Subset: IM |
Affiliation:
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Dept. of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Binding Sites / genetics Cell Line, Tumor Chondrocytes / metabolism* Collagen Type II / biosynthesis, genetics* DNA-Binding Proteins / antagonists & inhibitors*, metabolism Enhancer Elements, Genetic / genetics* High Mobility Group Proteins / genetics, metabolism Interleukin-1 / pharmacology Mice Molecular Mimicry Molecular Sequence Data Oligodeoxyribonucleotides / genetics* Promoter Regions, Genetic / genetics RNA, Messenger / analysis, metabolism SOX9 Transcription Factor Transcription Factors / antagonists & inhibitors*, genetics, metabolism Transcriptional Activation* / drug effects Transfection |
| Chemical | |
Reg. No./Substance:
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0/Col2a1 protein, mouse; 0/Collagen Type II; 0/DNA-Binding Proteins; 0/High Mobility Group Proteins; 0/Hivep1 protein, mouse; 0/Interleukin-1; 0/Oligodeoxyribonucleotides; 0/RNA, Messenger; 0/SOX9 Transcription Factor; 0/Sox9 protein, mouse; 0/Transcription Factors |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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