Characterization of MyD118, Gadd45, and proliferating cell nuclear antigen (PCNA) interacting domains - PCNA impedes MyD118 and Gadd45-mediated negative growth control
Characterization of MyD118, Gadd45, and proliferating cell nuclear antigen (PCNA) interacting domains - PCNA impedes MyD118 and Gadd45-mediated negative growth control
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DOI：
10.1074/jbc.275.22.16810
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发表时间：
2000-06-02
影响因子：
4.8
通讯作者：
Liebermann, DA
中科院分区：
文献类型：
--
作者：
Vairapandi, M;Azam, N;Liebermann, DA
MyD 118和Gadd 45是编码在负生长控制（包括生长抑制和凋亡）中起重要作用的蛋白质的相关基因。MyD 118和Gadd 45是先前显示与增殖细胞核抗原（PCNA）相互作用的相关蛋白，涉及DNA复制、DNA修复和细胞周期进程。为了确定MyD 118和Gadd 45与PCNA相互作用的作用，在这项工作中，我们试图确定相互作用的结构域，并分析这种相互作用在负生长控制中的意义。使用互补的体内和体外相互作用测定，PCNA的N-末端（1-46）和中间（100-127）区域被鉴定为含有MyD 118和Gadd 45相互作用结构域，而MyD 118和Gadd 45内的PCNA相互作用结构域定位于这些蛋白质的C末端（分别为氨基酸114-156和131-165）。这些发现提供了第一个证据，MyD 118和Gadd 45内的类似结构域介导与PCNA的相互作用。重要的是，异位表达的MyD 118或Gadd 45 N-末端肽，缺乏PCNA相互作用域，被发现抑制集落形成或诱导细胞凋亡比全长蛋白质更有效。这些发现表明，MyD 118或Gadd 45与PCNA的相互作用，在本质上，起到阻碍负生长控制的作用。
MyD118 and Gadd45 are related genes encoding for proteins that play important roles in negative growth control, including growth suppression and apoptosis. MyD118 and Gadd45 are related proteins that previously were shown to interact with proliferating cell nuclear antigen (PCNA), implicated in DNA replication, DNA repair, and cell cycle progression. To establish the role of MyD118 and Gadd45 interactions with PCNA, in this work we sought to identify the interacting domains and analyze the significance of this interaction in negative growth control. Using complementary in vivo and in vitro interaction assays the N-terminal (1-46) and middle (100-127) regions of PCNA were identified as harboring MyD118- and Gadd45 interacting domains, whereas PCNA interacting domains within MyD118 and Gadd45 were localized to the C termini of these proteins (amino acids 114-156 and 131-165, respectively). These findings provide first evidence that similar domains within MyD118 and Gadd45 mediate interactions with PCNA. Importantly, ectopic expression of MyD118 or Gadd45 N-terminal peptides, lacking the PCNA interacting domain, was found to suppress colony formation or induce apoptosis more efficiently than the full-length proteins. These findings suggest that interaction of MyD118 or Gadd45 with PCNA, in essence, serves to impede negative growth control.