| Project/Area Number |
12480210
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | Kobe University |
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Principal Investigator |
SAKAMOTO Hiroshi Kobe University, Faculty of Science, Professor, 理学部, 教授 (00187048)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 2001: ¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 2000: ¥7,900,000 (Direct Cost: ¥7,900,000)
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| Keywords | Hu protein / RNA-binding protein / neuron / protein-protein interaction / AU-rich element / 細胞内局在 / RNA結合ドメイン / 神経分化 / 線虫変異体 / RNA結合特異性 / 免疫沈降法 |
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| Research Abstract |
Hu proteins are RNA-binding proteins that are the vertebrate homologues of Drosophila ELAV, and are implicate in stabilization or enhanced translation of specific mRNAs with AU-rich elements (AREs) in the 3-untranslated region. Here, using the yeast two-hybrid system, we show that neuron-specific Hu proteins can interact with themselves. Immunoprecipitation assays demonstrated that the interaction between Hu proteins occurs in mammalian cells and is strongly enhanced in the presence of cellular RNA, Furthermore, using in situ chemical crosslinking assays, we found that HuD, one of the neuron-specific Hu proteins, multimerizes in cells. The crosslinked HuD multimers retain specific RNA-binding ability and can interact with additional Hu proteins. Consistent with this biochemical property, HuD showed granular distribution in two neurogenic cell lines. These results suggest that the RNA-bound form of HuD multimerizes cooperatively to form a specific granular structure that may serve as a site of post-transcriptional regulation of ARE-containing mRNAs.
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