Regulation by post-translation modifications in response to stress

通过翻译后修饰来应对压力的调节

• 批准号: 10801759
• 负责人: David Paul Toczyski
• 金额: $ 7.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10801759
• 关键词: Affect; Apoptotic; Area; BCL2L11 gene; Biological; Biological Process; Biology; CRISPR screen; CUL5 gene; Cells; Complex; Deubiquitinating Enzyme; Enzymes; Event; Gene Mutation; Human; Ligase; Messenger RNA; Mitochondria; Mutation; Nuclear Export; Pathway interactions; Post-Translational Protein Processing; Post-Translational Regulation; Protein Export Pathway; Proteins; Proteome; Regulation; Resistance; Set protein; Specificity; Stress; Ubiquitin; Ubiquitination; inhibitor; mRNA Export; member; response; ubiquitin isopeptidase; ubiquitin ligase

Project Summary/Abstract Ubiquitin ligases (E3s) represent a diverse and conserved group of enzymes, with over 600 members. These ligases collectively attach the small protein ubiquitin to more than twenty five percent of the proteome, thereby regulating the stability or activity of each target. Despite the importance of this set of enzymes, only a small percentage of ubiquitin ligases have well-characterized biological functions. We have conducted a CRISPR screen examining the sensitivity of mutations of genes encoding human ubiquitin ligases and deubiquitinating enzymes to a panel of inhibitors covering a broad range of biological pathways. From this screen, we identified a CUL5 specificity subunit, called WSB2, whose mutation renders cells sensitive to inhibitors of nuclear export and mitochondrial function. Preliminary studies identified two sets of proteins whose levels are affected by WSB2: the apoptotic factor BIM and the mRNA export proteins SNIP1 and THRAP3 (two members of the SNARP complex). In this application, we will determine whether these are independent substrates of the CUL5-WSB2 complex, or alternatively, whether targeting of SNIP1/THRAP3 affects BIM mRNA levels or localization. If SNIP1 and/or THRAP3 are direct substrates, we will examine the regulation of this ubiquitination event.

项目摘要/摘要 泛素连接酶(E3)是一类种类繁多、结构保守的酶，有600多个成员。这些 连接酶共同地将小蛋白泛素连接到超过25%的蛋白质组上，从而 调节每个靶标的稳定性或活性。尽管这套酶很重要，但只有一小部分 百分比的泛素连接酶具有良好的生物学功能。我们进行了一次CRISPR 人泛素连接酶基因突变与脱泛素化敏感性的筛查 从酶到一组抑制剂，覆盖了广泛的生物途径。从这个屏幕上，我们识别出 一种称为WSB2的CUL5特异性亚基，其突变使细胞对核出口抑制剂敏感 和线粒体功能。初步研究确定了两组蛋白质，其水平受 WSB2：凋亡因子BIM和信使核糖核酸输出蛋白SNIP1和THRAP3(SNARP的两个成员 复杂)。在本申请中，我们将确定这些是否为CUL5-WSB2的独立底物 复杂的，或者，靶向SNIP1/THRAP3是否影响BIM mRNA水平或定位。如果 SNIP1和/或THRAP3是直接底物，我们将研究这种泛素化事件的调节。

项目成果

Parallel Parkin: Cdc20 Takes a New Partner.

并行 Parkin：Cdc20 有了新合作伙伴。

• DOI: 10.1016/j.molcel.2015.09.015
• 发表时间: 2015
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Meza-Gutierrez,Fernando;Hundley,FrancesV;Toczyski,DavidP
• 通讯作者: Toczyski,DavidP

Isolation of ubiquitinated substrates by tandem affinity purification of E3 ligase-polyubiquitin-binding domain fusions (ligase traps).

通过将E3连接酶 - 聚氨酯结合结构域融合（连接酶陷阱）的串联纯度纯化（连接酶陷阱）分离出泛素化的底物。

• DOI: 10.1038/nprot.2016.008
• 发表时间: 2016-02
• 期刊: Nature protocols
• 影响因子: 14.8
• 作者: Mark KG;Loveless TB;Toczyski DP
• 通讯作者: Toczyski DP

The Yeast DNA Damage Checkpoint Kinase Rad53 Targets the Exoribonuclease, Xrn1.

酵母 DNA 损伤检查点激酶 Rad53 靶向外切核糖核酸酶 Xrn1。

• DOI: 10.1534/g3.118.200767
• 发表时间: 2018
• 期刊: G3 (Bethesda, Md.)
• 作者: Lao,JessicaP;Ulrich,KatieM;Johnson,JeffreyR;Newton,BillyW;Vashisht,AjayA;Wohlschlegel,JamesA;Krogan,NevanJ;Toczyski,DavidP
• 通讯作者: Toczyski,DavidP

Redundant targeting of Isr1 by two CDKs in mitotic cells.

• DOI: 10.1007/s00294-020-01110-x
• 发表时间: 2021-03
• 期刊: Current genetics
• 影响因子: 2.5
• 作者: Alme EB;Toczyski DP
• 通讯作者: Toczyski DP

Shelterin and subtelomeric DNA sequences control nucleosome maintenance and genome stability

• DOI: 10.15252/embr.201847181
• 发表时间: 2018-08
• 期刊: bioRxiv
• 作者: Thomas van Emden;Marta Forn;I. Forné;Zsuzsa Sarkadi;Matías Capella;Lucía Martín Caballero;Sabine Fischer-Burkart;Cornelia Brönner;M. Simonetta;D. Toczyski;M. Halić;A. Imhof;Sigurd Braun