MRNA EXPORT FROM THE NUCLEUS OF SACCHAROMYCES CEREVISIAE

酿酒酵母细胞核的 mRNA 输出

• 批准号: 3306029
• 负责人: ALAN Michael TARTAKOFF
• 金额: $ 16.59万
• 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/3306029
• 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; nucleic acid sequence; 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.

我们的目标是鉴定可用的温度敏感突变体 酿酒酵母(amn突变体)在酵母菌中积累PolA+mRNA 原子核温度为37℃。这些突变体分为11个互补组。 在确定Polya+RNA积累的程度后，可逆性 在37度时，mRNA的周转率，上位式 AMN的三个表型可区分类之间的关系 突变体，蛋白质在37度时进入细胞核的程度，以及 我们将研究37度时核包膜的形态变化。 使用野生型和37度的AMN细胞，5‘和3’的结构 然后将描述平均信使核糖核酸的末端，以及评估 剪接和正常剪接的前信使核糖核酸的命运。 此外，还将确定mRNA在核内的位置，并设置 将确定与核内mRNA相关的蛋白质。 进一步的研究将监测tRNA和rRNA的合成和去向 37度。同时，负责ts损伤的基因将被克隆。 通过互补和排序，Under和Under的后果 AMN基因的过度表达将被研究，AMN转录本将被 将针对相应的基因产生特征性和抗血清 产品。抗血清将用于定位AMN基因产物和 研究它们的生物合成和周转。

项目成果

A conditional yeast mutant deficient in mRNA transport from nucleus to cytoplasm.

一种条件性酵母突变体，其 mRNA 从细胞核转运至细胞质的能力存在缺陷。

• DOI: 10.1073/pnas.89.6.2312
• 发表时间: 1992
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Kadowaki,T;Zhao,Y;Tartakoff,AM
• 通讯作者: Tartakoff,AM

mRNA transport in yeast: time to reinvestigate the functions of the nucleolus.

酵母中的 mRNA 运输：是时候重新研究核仁的功能了。

• DOI: 10.1091/mbc.6.4.357
• 发表时间: 1995
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Schneiter,R;Kadowaki,T;Tartakoff,AM
• 通讯作者: Tartakoff,AM

Essential domains of the PRP21 splicing factor are implicated in the binding to PRP9 and PRP11 proteins and are conserved through evolution.

PRP21 剪接因子的重要结构域与 PRP9 和 PRP11 蛋白的结合有关，并且在进化过程中得到保留。

• 发表时间: 1996
• 期刊: RNA (New York, N.Y.)
• 作者: Rain,JC;Tartakoff,AM;Krämer,A;Legrain,P
• 通讯作者: Legrain,P

A yeast protein that bidirectionally affects nucleocytoplasmic transport.

一种双向影响核细胞质运输的酵母蛋白。

• DOI: 10.1242/jcs.108.1.265
• 发表时间: 1995
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Singleton,DR;Chen,S;Hitomi,M;Kumagai,C;Tartakoff,AM
• 通讯作者: Tartakoff,AM