| RRC ID |
3858
|
| 著者 |
Ikeda M, Inoue S, Muramatsu M, Minatogawa Y.
|
| タイトル |
Characterization and identification of a steroid receptor-binding protein, SRB-RGS.
|
| ジャーナル |
Biol Pharm Bull
|
| Abstract |
We cloned the cDNA of a novel steroid receptor-binding protein, SRB-RGS, which suppressed the estrogen receptor (ER)alpha-mediated and other promoter-driven transcriptional activities. This study revealed the interaction between the full-length SRB-RGS and full-length ERalpha or ERbeta by a coimmunoprecipitation assay. The full-length SRB-RGS and full-length ERalpha interacted in COS-7 cell by a mammalian two-hybrid system. The interaction between intrinsic SRB-RGS and ERs in the nuclear ER extract from the rat uteri was observed by the gel-shift assay. These results strongly suggested that SRB-RGS interacts with ERs bound to DNA (estrogen response element) in the nuclei of the cells. SRB-RGS suppressed very efficiently the ERalpha-, ERbeta-, and ERalpha+ERbeta-mediated transcriptional activities. Green fluorescence of enhanced green fluorescence protein (EGFP)-tagged SRB-RGS was localized both in the nucleus and in the cytoplasm. Intrinsic SRB-RGS was immunostained in the nucleus and the cytoplasm of HeLa cells. The putative SRB-RGS deduced from cDNA sequence was identified by the immunostaining and Western blotting by using the anti-SRB-RGS antibody. Overexpression of SRB-RGS induced the cell death in the HeLa cells. The nucleotide sequence of SRB-RGS cDNA that we cloned previously is identical with that of the newly isolated RGS3 cDNA. SRB-RGS could interact with ERs bound DNA in the nuclei of the cells and suppressed the ERs-mediated transcriptional activities.
|
| 巻・号 |
30(6)
|
| ページ |
1056-64
|
| 公開日 |
2007-6-1
|
| DOI |
10.1248/bpb.30.1056
|
| PII |
JST.JSTAGE/bpb/30.1056
|
| PMID |
17541154
|
| MeSH |
Animals
Base Sequence
Blotting, Western
COS Cells
Cell Death
Cell Nucleus / metabolism
Chloramphenicol O-Acetyltransferase / analysis
Chloramphenicol O-Acetyltransferase / metabolism
Chlorocebus aethiops
Cloning, Molecular
Cytoplasm / metabolism
DNA, Complementary / chemistry
DNA, Complementary / metabolism
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Electrophoresis, Polyacrylamide Gel
Estrogen Receptor alpha / genetics
Estrogen Receptor alpha / metabolism
Estrogen Receptor beta / genetics
Estrogen Receptor beta / metabolism
Female
Fluorescent Antibody Technique, Direct
Green Fluorescent Proteins / metabolism
HeLa Cells
Humans
Immunohistochemistry
Immunoprecipitation
Microscopy, Confocal
Protein Binding
RGS Proteins
Rats
Rats, Sprague-Dawley
Receptors, Estrogen / genetics*
Receptors, Estrogen / metabolism*
Receptors, Steroid / chemistry
Receptors, Steroid / genetics
Receptors, Steroid / metabolism*
Repressor Proteins / chemistry*
Repressor Proteins / genetics
Repressor Proteins / metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic
Two-Hybrid System Techniques
|
| IF |
1.863
|
| 引用数 |
4
|
|
WOS 分野
|
PHARMACOLOGY & PHARMACY
|
| リソース情報 |
| ヒト・動物細胞 |
COS-7(RCB0539)
HeLa(RCB0007) |
