RRC ID |
42234
|
著者 |
Yamagiwa H, Yamada Y, Bolander ME, Sarkar G.
|
タイトル |
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.
|
ジャーナル |
Mol Biotechnol
|
Abstract |
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.
|
巻・号 |
28(1)
|
ページ |
1-8
|
公開日 |
2004-9-1
|
DOI |
10.1385/MB:28:1:01
|
PII |
MB:28:1:01
|
PMID |
15456958
|
MeSH |
Animals
Base Sequence
Binding Sites / genetics
Cell Line, Tumor
Chondrocytes / metabolism*
Collagen Type II / biosynthesis
Collagen Type II / genetics*
DNA-Binding Proteins / antagonists & inhibitors*
DNA-Binding Proteins / metabolism
Enhancer Elements, Genetic / genetics*
High Mobility Group Proteins / genetics
High Mobility Group Proteins / metabolism
Interleukin-1 / pharmacology
Mice
Molecular Mimicry
Molecular Sequence Data
Oligodeoxyribonucleotides / genetics*
Promoter Regions, Genetic / genetics
RNA, Messenger / analysis
RNA, Messenger / metabolism
SOX9 Transcription Factor
Transcription Factors / antagonists & inhibitors*
Transcription Factors / genetics
Transcription Factors / metabolism
Transcriptional Activation* / drug effects
Transfection
|
IF |
2.022
|
引用数 |
3
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
ATDC5(RCB0565) |