The TOG protein Stu2/XMAP215 interacts covalently and noncovalently with SUMO.
The TOG protein Stu2/XMAP215 interacts covalently and noncovalently with SUMO.
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DOI：
10.1002/cm.21449
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发表时间：
2018-07
期刊：
影响因子：
--
通讯作者：
Miller RK
中科院分区：
文献类型：
--
作者：
Greenlee M;Alonso A;Rahman M;Meednu N;Davis K;Tabb V;Cook R;Miller RK
Stu 2 p是XMAP 215/Dis 1/ch-TOG家族微管相关蛋白的酵母成员，可促进微管聚合。然而，调节其活性的因素并不清楚。在这里，我们报告说，Stu 2 p在芽殖酵母酿酒酵母SUMO相互作用的共价和非共价机制。Stu 2 p通过双杂交分析与酵母SUMO Smt 3 p、其E2 Ubc 9 p和E3 Nfi 1 p相互作用。Stu 2 p中含有二聚化结构域的区域对于与SUMO和Ubc 9 p的相互作用是必要且充分的。发现Stu 2 p在体外和体内都被sumoylated。Stu 2 p在下拉试验中与SUMO共纯化，反之亦然。Stu 2 p还结合到SUMO的非缀合形式，表明Stu 2 p可以与SUMO非共价相互作用。此外，Stu 2 p与STUbL酶Ris 1 p相互作用。Stu 2 p还在下拉测定中与泛素共纯化，表明它可以被SUMO和泛素修饰。微管蛋白，一个主要的结合伙伴的Stu 2 p，也与SUMO非共价相互作用。通过双杂交分析，β-微管蛋白Tub 2 p与SUMO相互作用独立于微管应激子苯菌灵。总之，这些发现提高了由Stu 2 p介导的微管聚合活性通过sumoylation途径调节的可能性。
Stu2p is the yeast member of the XMAP215/Dis1/ch‐TOG family of microtubule‐associated proteins that promote microtubule polymerization. However, the factors that regulate its activity are not clearly understood. Here we report that Stu2p in the budding yeast Saccharomyces cerevisiae interacts with SUMO by covalent and noncovalent mechanisms. Stu2p interacted by two‐hybrid analysis with the yeast SUMO Smt3p, its E2 Ubc9p, and the E3 Nfi1p. A region of Stu2p containing the dimerization domain was both necessary and sufficient for interaction with SUMO and Ubc9p. Stu2p was found to be sumoylated both in vitro and in vivo. Stu2p copurified with SUMO in a pull‐down assay and vice versa. Stu2p also bound to a nonconjugatable form of SUMO, suggesting that Stu2p can interact noncovalently with SUMO. In addition, Stu2p interacted with the STUbL enzyme Ris1p. Stu2p also copurified with ubiquitin in a pull‐down assay, suggesting that it can be modified by both SUMO and ubiquitin. Tubulin, a major binding partner of Stu2p, also interacted noncovalently with SUMO. By two‐hybrid analysis, the beta‐tubulin Tub2p interacted with SUMO independently of the microtubule stressor, benomyl. Together, these findings raise the possibility that the microtubule polymerization activities mediated by Stu2p are regulated through sumoylation pathways.
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影响因子：
2.9
作者：
Alonso, Annabel;Greenlee, Matt;Matts, Jessica;Kline, Jake;Davis, Kayla J.;Miller, Rita K.
通讯作者：
Miller, Rita K.
影响因子：
64.5
作者：
Johnson, ES;Gupta, AA
通讯作者：
Gupta, AA
影响因子：
5.4
作者：
Elmore ZC;Donaher M;Matson BC;Murphy H;Westerbeck JW;Kerscher O
通讯作者：
Kerscher O
影响因子：
21.3
作者：
Bloom, K
通讯作者：
Bloom, K
影响因子：
64.5
作者：
Brouhard, Gary J.;Stear, Jeffrey H.;Hyman, Anthony A.
通讯作者：
Hyman, Anthony A.