CR6-interacting factor 1 interacts with Gadd45 family proteins and modulates the cell cycle
CR6-interacting factor 1 interacts with Gadd45 family proteins and modulates the cell cycle
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DOI：
10.1074/jbc.m212835200
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发表时间：
2003-07-25
影响因子：
4.8
通讯作者：
Shong, MH
中科院分区：
文献类型：
--
作者：
Chung, HK;Yi, YW;Shong, MH
Gadd45 蛋白家族包括 Gadd45alpha、MyD118/Gadd45beta 和 CR6/OIG37/Gadd45gamma。这些蛋白质在维持基因组稳定性和调节细胞周期方面发挥着重要作用。这项研究报告了一种名为 CR6 相互作用因子 1 (CRIF1) 的新型蛋白质的克隆，该蛋白质与 Gadd45alpha、MyD118/Gadd45beta 和 CR6/OIG37/Gadd45gamma 相互作用。通过体外谷胱甘肽 S-转移酶 Pull-down 测定和体内哺乳动物细胞双杂交测定以及免疫共沉淀测定确定 CRIF1 特异性结合 Gadd45 家族蛋白。 CRIF1 mRNA 在甲状腺、心脏、淋巴结、气管和肾上腺组织中高表达。 CRIF1 仅定位于细胞核并与 Gadd45gamma 共定位。重组CRIF1抑制免疫沉淀的Cdc2-cyclin B1和Cdk2-cyclin E的组蛋白H1激酶活性，并且抑制作用与Gadd45蛋白相加。 CRIF1 的过表达会增加 G(1) 期细胞的百分比，降低 S 期细胞的百分比，并抑制 NIH3T3 细胞的生长。通过转染小干扰RNA双链体下调内源性CRIF1，导致Rb磷酸化失活，并减少G(1)期细胞群。 CRIF1 的表达在肾上腺腺瘤和甲状腺乳头状癌中几乎检测不到，并且远低于邻近正常组织。本文的结果表明，CRIF1 是一种新型核蛋白，可与 Gadd45 相互作用，并可能在细胞周期进程和细胞生长的负调控中发挥作用。
The Gadd45 family of proteins includes Gadd45alpha, MyD118/Gadd45beta, and CR6/OIG37/Gadd45gamma. These proteins play important roles in maintaining genomic stability and in regulating the cell cycle. This study reports the cloning of a novel protein called CR6-interacting factor 1 (CRIF1) which interacts with Gadd45alpha, MyD118/Gadd45beta, and CR6/OIG37/Gadd45gamma. CRIF1 binds specifically to the Gadd45 family proteins, as determined by an in vitro glutathione S-transferase pull-down assay and an in vivo mammalian cell two-hybrid assay along with coimmunoprecipitation assays. CRIF1 mRNA is highly expressed in the thyroid gland, heart, lymph nodes, trachea, and adrenal tissues. CRIF1 localizes exclusively to the nucleus and colocalizes with Gadd45gamma. Recombinant CRIF1 inhibits the histone H1 kinase activity of immunoprecipitated Cdc2-cyclin B1 and Cdk2-cyclin E, and the inhibitory effects were additive with Gadd45 proteins. Overexpression of CRIF1 increases the percentage of cells in G(1), decreases the percentage of cells in S phase, and suppresses growth in NIH3T3 cells. The downregulation of endogenous CRIF1 by the transfection of the small interfering RNA duplexes resulted in the inactivation of Rb by phosphorylation and decreased the G(1) phase cell populations. Expression of CRIF1 is barely detectable in adrenal adenoma and papillary thyroid cancer and much lower than in adjacent normal tissue. The results presented here suggest that CRIF1 is a novel nuclear protein that interacts with Gadd45 and may play a role in negative regulation of cell cycle progression and cell growth.