REGULATION OF HISTIDINE BIOSYNTHESIS IN YEAST

酵母中组氨酸生物合成的调控

• 批准号: 2177702
• 负责人: GERALD R FINK
• 金额: $ 43.31万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2177702
• 关键词: Saccharomyces; aminoacid biosynthesis; cell cycle; cell growth regulation; chemical binding; chromosome aberrations; chromosome disorders; enzyme mechanism; enzyme substrate complex; eukaryote; fungal genetics; gene expression; gene induction /repression; genetic manipulation; genetic mapping; genetic promoter element; genetic recombination; genetic transcription; genetic translation; histidine; laboratory rabbit; molecular cloning; molecular genetics; mutagen testing; nucleic acid hybridization; nucleic acid sequence; proteins; regulatory gene; site directed mutagenesis; structural genes; synthetic nucleic acid; temperature sensitive mutant; transfer RNA; transposon /insertion element

Our goal is to understand the regulation of gene activity in the yeast Saccharomyces cerevisiae. The studies will focus on histidine biosynthesis as well as the transposable genetic element, Ty1. Both cis and trans-acting elements affecting expression of the histidine genes will be studied using fusions of regulatory sequences to E. coli Beta-galactosidase. The cis-acting sequences within the promoter will be dissected by a combination of site-directed mutagenesis and oligonucleotide synthesis. Various synthetic oligonucleotides will be substituted both at HIS4 and CYC1 to separate the promoter elements required for regulation, initiation, and maintenance of the basal level of transcription. Each of these promoter segments will be used as probes (both genetic and biochemical) for the trans-acting elements that interact with each of the cis-acting elements. In addition, genetic and biochemical analysis of the components required for permeation and cellular localization of histidine will be carried out. The mechanisms by which Ty elements transpose and activate genes can now be dissected. Again, both cis and trans-acting elements will be studied. The genes required in trans for transposition will be cloned and their role in the transposition process determined. The intermediates in transposition as well as the enzymes responsible for transposition will be isolated. Since the Ty element transposes through an RNA intermediate, we will try to isolate and characterize both the reverse transcriptase and RNase's involved in reverse transcription. Our assay system permits the analysis of the structural features of the Ty element that are required for transposition. By making mutations of various modified elements important cis-acting sequences will be identified: The ends of the element, the polypurine stretch, the polymerase binding site. Furthermore, mutations in the element should permit the isolation of intermediates in transposition and ultimately the reconstruction of the pathway of transposition.

我们的目标是了解酵母中基因活性的调节 酿酒酵母 这些研究将集中在组氨酸的生物合成 以及转座基因元件Ty 1。 顺式和 影响组氨酸基因表达的反式作用元件将被 研究使用调节序列与E.杆菌 β-半乳糖苷酶。 启动子内的顺式作用序列将是 通过定点诱变和寡核苷酸的组合来解剖 合成. 各种合成的寡核苷酸将被取代， HIS 4和CYC 1分离调控所需的启动子元件， 启动和维持转录的基础水平。 中的每 这些启动子片段将用作探针（遗传和 生物化学）的反式作用元件，与每一个相互作用， 顺式作用元件。 此外，还进行了遗传和生化分析， 组氨酸渗透和细胞定位所需的组分 将被执行。 Ty元件转座和激活基因的机制现在可以被解释为： 解剖。 同样，顺式和反式作用元件都将被研究。 的 转座所需的反式基因将被克隆，它们在转座中的作用将被证实。 转座过程确定。 转座中的中间体 以及负责转座的酶将被分离。 由于Ty元件通过RNA中间体转座，我们将尝试 分离并鉴定逆转录酶和RNA酶 参与逆转录。 我们的分析系统允许分析 Ty元素的结构特征， 换位 通过使各种修饰元件的突变变得重要 将鉴定顺式作用序列：元件的末端， 多嘌呤延伸，聚合酶结合位点。 此外，突变 该元件应允许分离转座中的中间体 并最终重建转座途径。

项目成果

A synthetic HIS4 regulatory element confers general amino acid control on the cytochrome c gene (CYC1) of yeast.

合成的 HIS4 调节元件赋予酵母细胞色素 c 基因 (CYC1) 一般氨基酸控制。

• DOI: 10.1073/pnas.82.2.498
• 发表时间: 1985
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Hinnebusch,AG;Lucchini,G;Fink,GR
• 通讯作者: Fink,GR

Ty-mediated gene expression of the LYS2 and HIS4 genes of Saccharomyces cerevisiae is controlled by the same SPT genes.

• DOI: 10.1073/pnas.81.8.2431
• 发表时间: 1984-04
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: G. Simchen;F. Winston;C. Styles;G. Fink

Mutations in POL1 increase the mitotic instability of tandem inverted repeats in Saccharomyces cerevisiae.

POL1 的突变增加了酿酒酵母串联反向重复序列的有丝分裂不稳定性。

• DOI: 10.1093/genetics/134.1.43
• 发表时间: 1993
• 期刊: Genetics
• 影响因子: 3.3
• 作者: Ruskin,B;Fink,GR
• 通讯作者: Fink,GR

Genetic variation in Saccharomyces cerevisiae: circuit diversification in a signal transduction network.

• DOI: 10.1534/genetics.112.145573
• 发表时间: 2012-12
• 期刊: Genetics
• 影响因子: 3.3
• 作者: Chin BL;Ryan O;Lewitter F;Boone C;Fink GR
• 通讯作者: Fink GR

HTS1 encodes both the cytoplasmic and mitochondrial histidyl-tRNA synthetase of Saccharomyces cerevisiae: mutations alter the specificity of compartmentation.

HTS1 编码酿酒酵母的细胞质和线粒体组氨酰-tRNA 合成酶：突变改变了区室划分的特异性。

• DOI: 10.1093/genetics/132.4.987
• 发表时间: 1992
• 期刊: Genetics
• 影响因子: 3.3
• 作者: Chiu,MI;Mason,TL;Fink,GR
• 通讯作者: Fink,GR