AKT介导的磷酸化增强ZNF322A的蛋白质稳定性和转录活性，从而促进肺癌的进展。,Oncogene

当前位置： X-MOL 学术 › Oncogene › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

AKT介导的磷酸化增强ZNF322A的蛋白质稳定性和转录活性，从而促进肺癌的进展。
Oncogene ( IF 6.9 ) Pub Date : 2019-08-09 , DOI: 10.1038/s41388-019-0928-x
Sheng-You Liao , I-Ying Kuo , Yu-Ting Chen , Pao-Chi Liao , Ya-Fen Liu , Hsin-Yi Wu , Wu-Wei Lai , Yi-Ching Wang

ZNF322A是致癌锌指转录因子。我们发表的结果表明，ZNF322A积极调节α-adducin（ADD1）和cyclin D1（CCND1）的转录，从而促进肺癌的致癌性。但是，ZNF322A蛋白功能的上游调节机制仍然难以捉摸。在这里，我们证明了AKT可以通过体外激酶测定和基于细胞的质谱分析使ZNF322A磷酸化。AKT的过表达促进了ZNF322A蛋白质的稳定性和转录活性，而AKT的敲低或用AKT抑制剂的作用抑制了这些作用。我们研究了AKT介导的磷酸化位点。Thr-150，Ser-224，Thr-234和Thr-262。AKT在Thr-262处的ZNF322A磷酸化促进了ZNF322A蛋白质的稳定性，因此增加了ADD1启动子的活性。有趣的是，Thr-150处的磷酸化 Ser-224和Thr-234增强了转录活性，而没有影响ZNF322A的蛋白质稳定性。染色质免疫沉淀和DNA亲和力沉淀试验表明，与野生型ZNF322A相比，ZNF322A磷酸化缺陷突变体Thr-150A，Ser-224A和Thr-234A减弱了染色质与ADD1和CCND1启动子的结合力和DNA结合亲和力。此外，AKT介导的Thr-150，Ser-224，Thr-234和Thr-262磷酸化促进了肺癌细胞在体外和体内的生长和转移。在临床上，来自150例肺癌患者的肿瘤标本中磷酸化ZNF322A（p-ZNF）的表达与活性磷酸化AKT（p-AKT）相关。多变量Cox回归分析表明，结合的p-AKT和p-ZNF表达谱是预测肺癌患者临床结局的独立因素。

"点击查看英文标题和摘要"

AKT-mediated phosphorylation enhances protein stability and transcription activity of ZNF322A to promote lung cancer progression.

ZNF322A is an oncogenic zinc-finger transcription factor. Our published results show that ZNF322A positively regulates transcription of alpha-adducin (ADD1) and cyclin D1 (CCND1) to promote tumorgenicity of lung cancer. However, the upstream regulatory mechanisms of ZNF322A protein function remain elusive. Here, we demonstrate that AKT could phosphorylate ZNF322A by in vitro kinase assay and cell-based mass spectrometry analysis. Overexpression of AKT promoted ZNF322A protein stability and transcriptional activity, whereas these effects were inhibited by knockdown of AKT or treating with AKT inhibitor. We studied AKT-mediated phosphorylation sites, viz. Thr-150, Ser-224, Thr-234, and Thr-262. ZNF322A phosphorylation at Thr-262 by AKT promoted ZNF322A protein stability thus increased ADD1 promoter activity. Interestingly, phosphorylation at Thr-150, Ser-224, and Thr-234 enhanced transcription activity without affecting protein stability of ZNF322A. Chromatin immunoprecipitation and DNA affinity precipitation assays showed that ZNF322A phosphorylation defective mutants Thr-150A, Ser-224A, and Thr-234A attenuated chromatin binding and DNA binding affinity to ADD1 and CCND1 promoters compared with wild-type ZNF322A. Furthermore, AKT-mediated Thr-150, Ser-224, Thr-234, and Thr-262 phosphorylation promoted lung cancer cell growth and metastasis in vitro and in vivo. Clinically, expression of phosphorylated ZNF322A (p-ZNF) correlated with actively phosphorylated AKT (p-AKT) in tumor specimens from 150 lung cancer patients. Multivariate Cox regression analysis indicated that combined p-AKT and p-ZNF expression profile was an independent factor to predict the clinical outcome in lung cancer patients. Our results reveal a new mechanism of AKT signaling in promoting ZNF322A protein stability and transcriptional activity in lung cancer cell, xenograft, and clinical models.

更新日期：2019-08-09

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南