Goodson M L, Park-Sarge O K, Sarge K D
Department of Biochemistry, Chandler Medical Center, University of Kentucky, Lexington 40536-0084, USA.
Mol Cell Biol. 1995 Oct;15(10):5288-93. doi: 10.1128/MCB.15.10.5288.
Heat shock factor 2 (HSF2) functions as a transcriptional regulator of heat shock protein gene expression in mammalian cells undergoing processes of differentiation and development. Our previous studies demonstrated high regulated expression and unusual constitutive DNA-binding activity of the HSF2 protein in mouse testes, suggesting that HSF2 functions to regulate heat shock protein gene expression in spermatogenic cells. The purpose of this study was to test whether HSF2 regulation in testes is associated with alterations in the HSF2 polypeptide expressed in testes relative to other mouse tissues. Our results show that mouse cells express not one but two distinct HSF2 proteins and that the levels of these HSF2 isoforms are regulated in a tissue-dependent manner. The testes express predominantly the 71-kDa HSF2-alpha isoform, while the heart and brain express primarily the 69-kDa HSF2-beta isoform. These isoforms are generated by alternative splicing of HSF2 pre-mRNA, which results in the inclusion of an 18-amino-acid coding sequence in the HSF2-alpha mRNA that is skipped in the HSF2-beta mRNA. HSF2 alternative splicing is also developmentally regulated, as our results reveal a switch in expression from the HSF2-beta mRNA isoform to the HSF2-alpha isoform during testis postnatal developmental. Transfection analysis shows that the HSF2-alpha protein, the predominant isoform expressed in testis cells, is a more potent transcriptional activator than the HSF2-beta isoform. These results reveal a new mechanism for the control of HSF2 function in mammalian cells, in which regulated alternative splicing is used to modulate HSF2 transcriptional activity in a tissue-dependent manner.
热休克因子2(HSF2)在经历分化和发育过程的哺乳动物细胞中作为热休克蛋白基因表达的转录调节因子发挥作用。我们之前的研究表明,HSF2蛋白在小鼠睾丸中具有高度调控的表达和异常的组成型DNA结合活性,这表明HSF2在生精细胞中发挥调节热休克蛋白基因表达的作用。本研究的目的是测试睾丸中HSF2的调节是否与睾丸中表达的HSF2多肽相对于其他小鼠组织的改变有关。我们的结果表明,小鼠细胞表达的不是一种而是两种不同的HSF2蛋白,并且这些HSF2同工型的水平以组织依赖性方式受到调节。睾丸主要表达71 kDa的HSF2-α同工型,而心脏和大脑主要表达69 kDa的HSF2-β同工型。这些同工型是由HSF2前体mRNA的可变剪接产生的,这导致HSF2-α mRNA中包含一个18个氨基酸的编码序列,而该序列在HSF2-β mRNA中被跳过。HSF2的可变剪接在发育过程中也受到调节,因为我们的结果显示,在睾丸出生后的发育过程中,表达从HSF2-β mRNA同工型转换为HSF2-α同工型。转染分析表明,睾丸细胞中表达的主要同工型HSF2-α蛋白比HSF2-β同工型是更强效的转录激活剂。这些结果揭示了哺乳动物细胞中控制HSF2功能的一种新机制,其中调控的可变剪接被用于以组织依赖性方式调节HSF2的转录活性。