論文 - 詳細
| RRC ID | 1412 |
|---|---|
| 著者 | den Engelsman J, Bennink EJ, Doerwald L, Onnekink C, Wunderink L, Andley UP, Kato K, de Jong WW, Boelens WC. |
| タイトル | Mimicking phosphorylation of the small heat-shock protein alphaB-crystallin recruits the F-box protein FBX4 to nuclear SC35 speckles. |
| ジャーナル | Eur J Biochem |
| Abstract |
The mammalian small heat shock protein alphaB-crystallin can be phosphorylated at three different sites, Ser19, Ser45 and Ser59. We compared the intracellular distribution of wild-type, nonphosphorylatable and all possible pseudophosphorylation mutants of alphaB-crystallin by immunoblot and immunocytochemical analyses of stable and transiently transfected cells. We observed that pseudophosphorylation at two (especially S19D/S45D) or all three (S19D/S45D/S59D) sites induced the partial translocation of alphaB-crystallin from the detergent-soluble to the detergent-insoluble fraction. Double immunofluorescence studies showed that the pseudophosphorylation mutants localized in nuclear speckles containing the splicing factor SC35. The alphaB-crystallin mutants in these speckles were resistant to mild detergent treatment, and also to DNase I or RNase A digestion, indicating a stable interaction with one or more speckle proteins, not dependent on intact DNA or RNA. We further found that FBX4, an adaptor protein of the ubiquitin-protein isopeptide ligase SKP1/CUL1/F-box known to interact with pseudophosphorylated alphaB-crystallin, was also recruited to SC35 speckles when cotransfected with the pseudophosphorylation mutants. Because SC35 speckles also react with an antibody against alphaB-crystallin endogenously phosphorylated at Ser45, our findings suggest that alphaB-crystallin has a phosphorylation-dependent role in the ubiquitination of a component of SC35 speckles. |
| 巻・号 | 271(21) |
| ページ | 4195-203 |
| 公開日 | 2004-11-1 |
| DOI | 10.1111/j.1432-1033.2004.04359.x |
| PII | EJB4359 |
| PMID | 15511225 |
| MeSH | Binding Sites Cell Nucleus / metabolism* Deoxyribonuclease I / metabolism Detergents / pharmacology Endopeptidases / metabolism F-Box Proteins / chemistry* HeLa Cells Humans Immunoblotting Immunohistochemistry Microscopy, Fluorescence Mutation Nuclear Proteins / metabolism* Phosphorylation Plasmids / metabolism Protein Binding Protein Transport Ribonuclease, Pancreatic / metabolism Ribonucleoproteins / metabolism* Serine / chemistry Serine-Arginine Splicing Factors Subcellular Fractions / metabolism Ubiquitin / metabolism alpha-Crystallin B Chain / chemistry* alpha-Crystallin B Chain / metabolism |
| 引用数 | 59 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | anti-αB-crystallin(RCB1304) |