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BIOGENESIS AND TRAFFICKING OF U3 SMALL NUCLEOLAR RNPS

BIOGENESIS AND TRAFFICKING OF U3 SMALL NUCLEOLAR RNPS
U3 小核仁 RNPS 的生物发生和运输
批准号：
6630683
负责人：
MICHAEL P TERNS
金额：
$6.5万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
项目状态：
已结题
起止时间：
1997-08-01 至 2003-04-09
来源：
https://reporter.nih.gov/project-details/6630683
关键词：
RNA binding protein Xenopus oocyte affinity chromatography complementary DNA crosslink electron microscopy genetic library intracellular transport light microscopy nucleic acid sequence nucleolus posttranscriptional RNA processing posttranslational modifications protein biosynthesis protein structure function protein transport ribosomal RNA ribosomal proteins small nuclear ribonucleoproteins

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中文摘要
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英文摘要
DESCRIPTION (adapted from application): The PI proposes to study all posttranscriptional steps involved in biogenesis of the U3 small nucleolar ribonucleoprotein particle (U3 snoRNP). These steps include binding of proteins to the precursor and mature forms of U3 snoRNA; hypermethylation of the 5' cap; nuclear retention of the RNA, RNP intermediates, and the mature snoRNP; and finally, relocalization of the U3 snoRNP from the nucleoplasm to the nucleolus. The experimental system of choice will be the Xenopus oocyte for all the usual reasons. In particular, microinjected U3 snoRNAs are reconstituted into U3 snoRNPs, and the subcellular localization of the resulting RNPs (cytoplasm, nucleoplasm, nucleoli) are easily determined both by biochemical fractionation and by traditional light microscopy. The Xenopus assay system will be used to determine the protein composition of reconstituted U3 snoRNPs, and the role of various U3 snoRNA sequences in protein binding, snoRNP reconstitution, and nucleolar localization. U3 snoRNP proteins will be identified by analytical assays (crosslinking, affinity selection) but cloned by two hybrid screens (for those that might interact with fibrillarin) and three hybrid screens (for those that might interact directly with U3 snoRNA).
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