Association of CAD, a multifunctional protein involved in pyrimidine synthesis, with mLST8, a component of the mTOR complexes.
Association of CAD, a multifunctional protein involved in pyrimidine synthesis, with mLST8, a component of the mTOR complexes.
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DOI：
10.1186/1423-0127-20-24
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发表时间：
2013-04-18
影响因子：
11
通讯作者：
Yonezawa K
中科院分区：
文献类型：
--
作者：
Nakashima A;Kawanishi I;Eguchi S;Yu EH;Eguchi S;Oshiro N;Yoshino K;Kikkawa U;Yonezawa K
mTOR是一种遗传上保守的丝氨酸/苏氨酸蛋白激酶，其控制细胞生长、增殖和存活。多功能蛋白CAD催化嘧啶从头合成的最初三个步骤，通过与不同蛋白激酶的磷酸化反应来调节，但与mTOR蛋白激酶的关系尚不清楚。从用FLAG-mLST 8载体转染的HEK 293细胞中回收CAD作为与mLST 8（mTOR复合物的组分）的结合蛋白。这两种蛋白质的缔合通过共免疫沉淀法确认，随后通过转染的myc-CAD和FLAG-mLST 8以及细胞中内源性蛋白质的免疫印迹分析确认。使用突变构建体的分析表明CAD具有多于一个与mLST 8结合的区域，并且mLST 8以不同的方式识别CAD和mTOR。CAD酶活性在耗尽氨基酸和血清的细胞中降低，其中mTOR活性被抑制。所获得的结果表明，mLST 8在CAD和mTOR之间起桥梁作用，并且在信号传导机制中起作用，其中CAD通过与mLST 8的结合在mTOR通路中被调节。
mTOR is a genetically conserved serine/threonine protein kinase, which controls cell growth, proliferation, and survival. A multifunctional protein CAD, catalyzing the initial three steps in de novo pyrimidine synthesis, is regulated by the phosphorylation reaction with different protein kinases, but the relationship with mTOR protein kinase has not been known. CAD was recovered as a binding protein with mLST8, a component of the mTOR complexes, from HEK293 cells transfected with the FLAG-mLST8 vector. Association of these two proteins was confirmed by the co-immuoprecipitaiton followed by immunoblot analysis of transfected myc-CAD and FLAG-mLST8 as well as that of the endogenous proteins in the cells. Analysis using mutant constructs suggested that CAD has more than one region for the binding with mLST8, and that mLST8 recognizes CAD and mTOR in distinct ways. The CAD enzymatic activity decreased in the cells depleted of amino acids and serum, in which the mTOR activity is suppressed. The results obtained indicate that mLST8 bridges between CAD and mTOR, and plays a role in the signaling mechanism where CAD is regulated in the mTOR pathway through the association with mLST8.
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影响因子：
3.9
作者：
Avruch, Joseph;Long, Xiaomeng;Kikkawa, Ushio
通讯作者：
Kikkawa, Ushio
影响因子：
16
作者：
Kim, DH;Sarbassov, DD;Sabatini, DM
通讯作者：
Sabatini, DM
影响因子：
4.3
作者：
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通讯作者：
Evans, David R.
影响因子：
2.1
作者：
Oshiro, N;Yoshino, K;Yonezawa, K
通讯作者：
Yonezawa, K
影响因子：
21.3
作者：
Jacinto, E;Loewith, R;Hall, MN
通讯作者：
Hall, MN