| Budget Amount *help |
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
We have cloned cDNAs encoding mHuD,mHuC and Mel-N1, the mouse homologues of a human nervous system-specific RNA-binding protein. Two major mRNA transcripts for each protein were detected predominantly in the adult mouse brain. To gain insight into the RNA binding specificity of these proteins, we performed iterative in vitro RNA selection. The resulting in vitro selected RNAs were found to contain the AU-rich element (ARE), which is involved in the regulation of mRNA stability. Our studies further reveal that Mel-N1 can bind to its own 3' untranslated region (3'UTR) as well as to the c-fos 3'UTR,and is localized predominantly in the cytoplasmic region in cells, suggesting that post-transcriptional autoregulation of Mel-N1 gene expression occurs in vivo. In addtion, we have found that two mHuC isoforms are generated by alternative splicing. Iterative in vitro RNA selection and binding analyzes showed that both HuC isoforms can bind with almost identical specificity to sequences similar to ARE.Functional domain mapping using mHuC deletion mutants showed that the first RRM binds to ARE,that the second RRM has no RNA-binding activity by itself, but facilitates ARE-binding by the first RRM,and that the third RRM has specific binding activity for the poly (A) sequence.
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