Functional domains of the Rsp5 ubiquitin-protein ligase.
Functional domains of the Rsp5 ubiquitin-protein ligase.
复制标题
Rsp5 泛素蛋白连接酶的功能域。
DOI：
10.1128/mcb.19.1.342
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发表时间：
1999
影响因子：
5.3
通讯作者：
Huibregtse,JM
中科院分区：
文献类型：
--
作者：
Wang,G;Yang,J;Huibregtse,JM
RSP5是酿酒酵母的必需基因，编码Hect结构域的E3泛素蛋白连接酶。Hect E3蛋白被认为由两个广泛的功能结构域组成：一个由大约350个氨基酸组成的保守的催化羧基末端结构域(Hect结构域)和一个包含底物特异性决定因素的大的非保守的氨基末端结构域。我们在这里报告了Rsp5在体内发挥基本功能所需的最小区域的图谱，与Rsp5底物(Rpb1，RNA聚合酶II的大亚基)稳定相互作用所需的最小区域，以及发现hect结构域本身就足以形成泛素-硫酯中间体。Hect结构域内的突变影响形成泛素-硫酸酯的能力或催化底物泛素化在体内的功能丧失，强烈表明Rsp5的泛素蛋白连接酶活性与其基本功能存在内在联系。Rsp5的氨基末端含有三个WW结构域和一个C2钙结合区。三个WW结构域中的两个是体内必需功能所必需的，而C2结构域不是，Rpb1结合和泛素化的要求存在于体内功能所需的区域内。综上所述，这些结果支持hect E3功能的双域模型，并表明WW结构域在识别Rsp5的至少部分底物方面发挥作用，包括与其基本功能相关的底物。此外，我们还发现，HUL4(YJR036C)和HUL5(YGL141W)这两个酵母的其他两个hect E3基因完全中断的单倍体酵母菌株是活的。
RSP5, an essential gene ofSaccharomyces cerevisiae, encodes a hect domain E3 ubiquitin-protein ligase. Hect E3 proteins have been proposed to consist of two broad functional domains: a conserved catalytic carboxyl-terminal domain of approximately 350 amino acids (the hect domain) and a large, nonconserved amino-terminal domain containing determinants of substrate specificity. We report here the mapping of the minimal region of Rsp5 necessary for its essential in vivo function, the minimal region necessary to stably interact with a substrate of Rsp5 (Rpb1, the large subunit of RNA polymerase II), and the finding that the hect domain, by itself, is sufficient for formation of the ubiquitin-thioester intermediate. Mutations within the hect domain that affect either the ability to form a ubiquitin-thioester or to catalyze substrate ubiquitination abrogate in vivo function, strongly suggesting that the ubiquitin-protein ligase activity of Rsp5 is intrinsically linked to its essential function. The amino-terminal region of Rsp5 contains three WW domains and a C2 calcium-binding domain. Two of the three WW domains are required for the essential in vivo function, while the C2 domain is not, and requirements for Rpb1 binding and ubiquitination lie within the region required for in vivo function. Together, these results support the two-domain model for hect E3 function and indicate that the WW domains play a role in the recognition of at least some of the substrates of Rsp5, including those related to its essential function. In addition, we show that haploid yeast strains bearing complete disruptions of either of two other hect E3 genes of yeast, designatedHUL4(YJR036C) andHUL5(YGL141W), are viable.
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影响因子：
3.6
作者：
HEIN, C;SPRINGAEL, JY;ANDRE, B
通讯作者：
ANDRE, B
影响因子：
11.4
作者：
HOFMANN, JFX;BEACH, D
通讯作者：
BEACH, D
影响因子：
2.5
作者：
Karagiannis, J;Saleki, R;Young, PG
通讯作者：
Young, PG
粟酒裂殖酵母细胞对营养饥饿和交配信息素信号的反应需要磷脂酰肌醇 3-磷酸 5-激酶
影响因子：
2.1
作者：
M. Morishita;Fusako Morimoto;K. Kitamura;T. Koga;Y. Fukui;H. Maekawa;I. Yamashita;C. Shimoda
通讯作者：
C. Shimoda
DOI：
10.1007/978-1-4615-7148-3
发表时间：
1976
期刊：
Molecular and General Genetics MGG
影响因子：
--
作者：
R. King
通讯作者：
R. King