MRNA EXPORT FROM THE NUCLEUS OF SACCHAROMYCES CEREVISIAE

酿酒酵母细胞核的 mRNA 输出

• 批准号: 3306028
• 负责人: ALAN Michael TARTAKOFF
• 金额: $ 15.91万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3306028
• 关键词: RNA biosynthesis; RNA splicing; SDS polyacrylamide gel electrophoresis; Saccharomyces; affinity chromatography; antibody; beta galactosidase; cell nucleus; electron microscopy; freeze etching; fungal genetics; fusion gene; gene complementation; gene expression; immunocytochemistry; immunofluorescence technique; immunoprecipitation; in situ hybridization; intracellular transport; laboratory rabbit; messenger RNA; molecular cloning; nuclear membrane; phosphorus; precursor mRNA; protein transport; protoplast /spheroplast; radionuclides; radiotracer; ribosomal RNA; temperature sensitive mutant; transfer RNA; tritium; western blottings

The goal is to characterize available temperature-sensitive mutants of Saccharomyces cerevisiae (amn mutants) which accumulate polyA+ mRNA in the nucleus at 37oC. These mutants fall into eleven complementation groups. After determining the extent of polyA+ RNA accumulation, the reversibility of mRNA accumulation, and the rate of turnover of mRNA at 37o, epistatic relations among three phenotypically-distinguishable classes of amn mutants, the extent of protein import into the nucleus at 37o, and morphological alterations of the nuclear envelope at 37o will be studied. Using wild type and amn cells at 37o, the structure of the 5' and 3' terminae of average mRNA will then be characterized, along with assessment of splicing and the fate of a pre-mRNA which normally is spliced. Moreover, mRNA intranuclear location will be determined and the set of proteins with which intranuclear mRNA is associated will be identified. Further studies will monitor the synthesis and fate of tRNA and rRNAs at 37o. In parallel, the genes responsible for the Ts- lesions will be cloned by complementation and sequenced, the consequences of under and overexpression of AMN genes will be studied, AMN transcripts will be characterized and antisera will be raised against the corresponding gene products. The antisera will be used to localize the AMN gene products and to study their biosynthesis and turnover.

目的是表征可用的温度敏感突变体 酿酒酵母（AMN突变体）在累积Polya+ mRNA 37oC的核。 这些突变体属于十一个互补组。 确定Polya+ RNA积累的程度后，可逆性 mRNA的积累和37o上mRNA的离职率 AMN的三个表型不同的类别之间的关系 突变体，蛋白质在37o时进口到细胞核的程度， 将研究37o时核包膜的形态改变。 在37O处使用野生型和AMN细胞，5'和3'的结构 然后将表征平均mRNA的末端以及评估 剪接和通常被剪接的前MRNA的命运。 此外，将确定mRNA核内位置，并集合 将发现与核内mRNA相关的蛋白质。 进一步的研究将监测tRNA和rRNA的合成和命运 37o。 同时，将克隆负责TS率的基因 通过互补和测序，下层的后果 将研究AMN基因的过表达，AMN转录本将是 特征和抗血清将针对相应的基因提出 产品。 抗血清将用于定位AMN基因产品，并且 研究他们的生物合成和周转率。