Stacey A Mattison, Gregory L Blatch, Adrienne L Edkins
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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.</p>","PeriodicalId":9812,"journal":{"name":"Cell Stress and Chaperones","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2017-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352595/pdf/","citationCount":"0","resultStr":"{\"title\":\"HOP expression is regulated by p53 and RAS and characteristic of a cancer gene signature.\",\"authors\":\"Stacey A Mattison, Gregory L Blatch, Adrienne L Edkins\",\"doi\":\"10.1007/s12192-016-0755-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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.</p>\",\"PeriodicalId\":9812,\"journal\":{\"name\":\"Cell Stress and Chaperones\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2017-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352595/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cell Stress and Chaperones\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1007/s12192-016-0755-8\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2016/12/16 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cell Stress and Chaperones","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/s12192-016-0755-8","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2016/12/16 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
Hsp70/Hsp90 组织蛋白(HOP)是 Hsp90 合子机器中客户蛋白从 Hsp70 转移到 Hsp90 所必需的辅助合子。虽然 HOP 在各种癌症中上调,但关于 HOP 在癌症中如何表达的体外研究信息却很有限。本研究的主要目的是确定 HOP 启动子并研究其在癌细胞中的活性。对 HOP 基因的 -2500 至 +16 bp 区域进行的生物信息学分析发现了一个大的 CpG 岛和一系列假定的顺式元件。许多顺式元件可能与转录因子结合,而转录因子会被致癌途径激活。荧光素酶报告实验表明,HOP 基因的上游区域包含一个体外活性启动子。对该区域的截断表明,HOP 核心启动子区域为 -855 至 +16 bp。在表达突变或非活性 p53 的 Hs578T、HEK293T 和 SV40 转化的 MEF1 细胞系中,HOP 启动子的活性最高。在突变 p53 背景下,表达野生型 p53 会导致启动子活性降低,而在 HeLa 细胞中抑制野生型 p53 会增加 HOP 启动子活性。此外,在含有非活性 p53 的 Hs578T 和 HEK293T 细胞系中,HRAS 的表达增加了 HOP 启动子的活性。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 的表达可能同时受到 RAS 激活和 p53 抑制的调控。总之,这些数据表明,HOP 可能是由突变 p53 和突变 RAS 共同诱导的癌症基因特征的一部分,与细胞转化有关。
HOP expression is regulated by p53 and RAS and characteristic of a cancer gene signature.
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.