Mattison Stacey A, Blatch Gregory L, Edkins Adrienne L
Biomedical Biotechnology Research Unit, Department of Biochemistry and Microbiology, Rhodes University, Grahamstown, South Africa.
Centre for Chronic Disease, College of Health and Biomedicine, Victoria University, Melbourne, Australia.
Cell Stress Chaperones. 2017 Mar;22(2):213-223. doi: 10.1007/s12192-016-0755-8. Epub 2016 Dec 16.
The Hsp70/Hsp90 organising protein (HOP) is a co-chaperone essential for client protein transfer from Hsp70 to Hsp90 within the Hsp90 chaperone machine. Although HOP is upregulated in various cancers, there is limited information from in vitro studies on how HOP expression is regulated in cancer. The main objective of this study was to identify the HOP promoter and investigate its activity in cancerous cells. Bioinformatic analysis of the -2500 to +16 bp region of the HOP gene identified a large CpG island and a range of putative cis-elements. Many of the cis-elements were potentially bound by transcription factors which are activated by oncogenic pathways. Luciferase reporter assays demonstrated that the upstream region of the HOP gene contains an active promoter in vitro. Truncation of this region suggested that the core HOP promoter region was -855 to +16 bp. HOP promoter activity was highest in Hs578T, HEK293T and SV40- transformed MEF1 cell lines which expressed mutant or inactive p53. In a mutant p53 background, expression of wild-type p53 led to a reduction in promoter activity, while inhibition of wild-type p53 in HeLa cells increased HOP promoter activity. Additionally, in Hs578T and HEK293T cell lines containing inactive p53, expression of HRAS increased HOP promoter activity. However, HRAS activation of the HOP promoter was inhibited by p53 overexpression. These findings suggest for the first time that HOP expression in cancer may be regulated by both RAS activation and p53 inhibition. Taken together, these data suggest that HOP may be part of the cancer gene signature induced by a combination of mutant p53 and mutated RAS that is associated with cellular transformation.
热休克蛋白70/热休克蛋白90组织蛋白(HOP)是一种辅助伴侣蛋白,对于热休克蛋白90伴侣机制中客户蛋白从热休克蛋白70转移至热休克蛋白90至关重要。尽管HOP在多种癌症中表达上调,但关于癌症中HOP表达如何调控的体外研究信息有限。本研究的主要目的是鉴定HOP启动子并研究其在癌细胞中的活性。对HOP基因-2500至+16 bp区域的生物信息学分析确定了一个大的CpG岛和一系列假定的顺式元件。许多顺式元件可能被致癌途径激活的转录因子所结合。荧光素酶报告基因检测表明,HOP基因的上游区域在体外含有一个活性启动子。该区域的截短表明HOP核心启动子区域为-855至+16 bp。HOP启动子活性在表达突变型或无活性p53的Hs578T、HEK293T和SV40转化的MEF1细胞系中最高。在突变型p53背景下,野生型p53的表达导致启动子活性降低,而在HeLa细胞中抑制野生型p53则增加HOP启动子活性。此外,在含有无活性p53的Hs578T和HEK293T细胞系中,HRAS的表达增加了HOP启动子活性。然而,p53过表达抑制了HRAS对HOP启动子的激活。这些发现首次表明,癌症中HOP的表达可能受RAS激活和p53抑制的双重调控。综上所述,这些数据表明,HOP可能是由与细胞转化相关的突变型p53和突变型RAS组合诱导的癌症基因特征的一部分。