Fu Guolong, Wei Youheng, Wang Xiaoli, Yu Long
a State Key Laboratory of Genetic Engineering, School of Life Sciences, Institute of Genetics , Fudan University , Shanghai , P.R. China.
b Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, School of Pharmaceutical Sciences , Wuhan University , Wuhan , P.R. China.
Biosci Biotechnol Biochem. 2016 Jun;80(6):1088-94. doi: 10.1080/09168451.2016.1146067. Epub 2016 Mar 4.
We aimed to investigate the internal existence status of testis-specific serine/threonine kinase 4 (Tssk4) and the interaction of Tssk4 and Cre-responsive element modulator (Crem). The internal existence status of Tssk4 in testis of mice was detected using western blotting and dephosphorylation method. The interaction of Tssk4 and Crem was analyzed by western blotting, immunohistochemistry, immunofluorescence, in vitro co-immunoprecipitation assays, and in vitro kinase assay. The results revealed that Tssk4 existed in testis both in phosphorylation and unphosphorylation status by a temporal manner with the development of testis. Immunofluorescence results showed that Tssk4 had identical distribution pattern with Crem in testis, which was utterly different to the localization of Cre-responsive element binding (Creb). In conclusion, our study demonstrated that phosphorylated Tssk4 might participate in testis genes expressions by phosphorylating Crem at Ser-117.
我们旨在研究睾丸特异性丝氨酸/苏氨酸激酶4（Tssk4）的内在存在状态以及Tssk4与Cre反应元件调节剂（Crem）的相互作用。采用蛋白质免疫印迹法和去磷酸化方法检测Tssk4在小鼠睾丸中的内在存在状态。通过蛋白质免疫印迹法、免疫组织化学、免疫荧光、体外共免疫沉淀试验和体外激酶试验分析Tssk4与Crem的相互作用。结果显示，随着睾丸的发育，Tssk4以时间依赖性方式在睾丸中以磷酸化和非磷酸化状态存在。免疫荧光结果表明，Tssk4在睾丸中的分布模式与Crem相同，这与Cre反应元件结合蛋白（Creb）的定位完全不同。总之，我们的研究表明，磷酸化的Tssk4可能通过在丝氨酸117位点磷酸化Crem来参与睾丸基因的表达。
