Yuan Xiaoqin, Kuramitsu Yasuhiro, Furumoto Hiroko, Zhang Xiulian, Hayashi Eiko, Fujimoto Masanori, Nakamura Kazuyuki
Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.
Electrophoresis. 2007 Jun;28(12):2018-26. doi: 10.1002/elps.200600821.
Heat stress causes severe constraints on numerous physiological functions of cells, such as the repression of splicing of mRNA precursors. In this study, we performed proteomic profiling of a nuclear fraction of Jurkat cells during heat stress using 2-DE and LC-MS/MS. We found 10 protein spots whose expression had changed after heat stress at 43 degrees C for 30 min. Seven of those protein spots, periodic tryptophan protein 1 homolog (PWP1), importin beta-1 subunit, sumoylated protein, splicing factor 3a subunit 3 (SF3a3), TAR DNA-binding protein 43, U2 small nuclear ribonucleoprotein auxiliary factor 35 kDa subunit (U2AF35) and small ubiquitin-related modifier-1 (SUMO-1) were downregulated, while three other protein spots, Protein SET, 40S ribosomal protein SA and 60S acidic ribosomal protein P0 were upregulated by the heat stress. We focused on the downregulation of two splicing factors, which might participate in the repression of pre-mRNA processing by heat stress, leading to cell apoptosis.
热应激对细胞的多种生理功能造成严重限制，例如抑制mRNA前体的剪接。在本研究中，我们利用二维电泳（2-DE）和液相色谱-串联质谱（LC-MS/MS）对热应激期间Jurkat细胞的核组分进行了蛋白质组分析。我们发现43℃热应激30分钟后有10个蛋白点的表达发生了变化。其中7个蛋白点，即周期性色氨酸蛋白1同源物（PWP1）、输入蛋白β-1亚基、SUMO化蛋白、剪接因子3a亚基3（SF3a3）、TAR DNA结合蛋白43、U2小核核糖核蛋白辅助因子35 kDa亚基（U2AF35）和小泛素相关修饰物1（SUMO-1）表达下调，而另外3个蛋白点，即蛋白SET、40S核糖体蛋白SA和60S酸性核糖体蛋白P0在热应激后表达上调。我们重点关注了两个剪接因子的下调，它们可能参与热应激对前体mRNA加工的抑制，从而导致细胞凋亡。
