Sykes K, Gething M J, Sambrook J
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235-9050.
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5853-7. doi: 10.1073/pnas.90.12.5853.
The cyclophilins (CYPs) and FK506 binding proteins (FKBPs) are two families of distinct proline isomerases that are targets for a number of clinically important immunosuppressive drugs. Members of both families catalyze cis/trans isomerization of peptidyl-prolyl bonds, which can be a rate-limiting step during protein folding in vitro and in vivo. We demonstrate in Saccharomyces cerevisiae that heat shock causes a 2- to 3-fold increase in the level of mRNA encoded by the major cytoplasmic CYP gene, CYP1. The cloned CYP1 promoter confers heat-inducible expression upon a reporter gene, and transcriptional induction is mediated through sequences similar to the consensus heat shock response element. Disruption of CYP1 decreases survival of cells following exposure to high temperatures, indicating that CYP1 plays a role in the stress response. A second CYP gene, CYP2, encodes a cyclophilin that is located within the secretory pathway. Its expression is also stimulated by heat shock, and cells containing a disrupted CYP2 allele are more sensitive than wild-type cells to heat. By contrast, expression of the FKB1 gene, which encodes a cytoplasmic member of the yeast FKBP family, is neither heat responsive nor necessary for survival after exposure to heat stress.
亲环蛋白(CYPs)和FK506结合蛋白(FKBPs)是两类不同的脯氨酸异构酶家族,它们是多种临床上重要的免疫抑制药物的作用靶点。这两个家族的成员都催化肽基 - 脯氨酰键的顺/反异构化,这在体外和体内蛋白质折叠过程中可能是一个限速步骤。我们在酿酒酵母中证明,热休克会使主要细胞质CYP基因CYP1编码的mRNA水平增加2至3倍。克隆的CYP1启动子赋予报告基因热诱导表达,并且转录诱导是通过与共有热休克反应元件相似的序列介导的。破坏CYP1会降低细胞在高温下的存活率,表明CYP1在应激反应中起作用。第二个CYP基因CYP2编码一种位于分泌途径中的亲环蛋白。它的表达也受热休克刺激,并且含有破坏的CYP2等位基因的细胞比野生型细胞对热更敏感。相比之下,编码酵母FKBP家族细胞质成员的FKB1基因的表达既不受热响应,也不是热应激后存活所必需的。