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HOP expression is regulated by p53 and RAS and characteristic of a cancer gene signature

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Cell Stress and Chaperones Aims and scope

Abstract

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.

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Acknowledgements

This research was supported by funding from the South African Research Chairs Initiative of the Department of Science and Technology (DST) and the National Research Foundation of South Africa (NRF) (Grant No. 98566), the Cancer Association of South Africa (CANSA), Medical Research Council South Africa (MRC-SA) with funds from the National Treasury under its Economic Competitiveness and Support Package and Rhodes University. SAM was the recipient of a postgraduate bursary from the NRF and German Academic Exchange Service (DAAD). The views expressed are those of the authors and should not be attributed to the DST, NRF, CANSA, MRC-SA or Rhodes University.

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  1. Biomedical Biotechnology Research Unit, Department of Biochemistry and Microbiology, Rhodes University, Grahamstown, South Africa

    Stacey A Mattison, Gregory L Blatch & Adrienne L Edkins

  2. Centre for Chronic Disease, College of Health and Biomedicine, Victoria University, Melbourne, Australia

    Gregory L Blatch

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  1. Stacey A Mattison
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Correspondence to Adrienne L Edkins.

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Supplementary Fig. 1

Confirmation of protein expression from plasmids. HeLa cells were transfected with pcDNA3-FLAG-p53 (a), pBABE-c-RAF 22 W (b), mEGFP-HRAS, mEGFP-HRAS G12 V, mEGFP-HRAS S17 N (c) or with water as a control. Whole cell lysates were produced 48 h after transfection and western blots were performed using 50 μg total protein with antibodies against FLAG, Raf1 and EGFP, respectively. (GIF 59 kb)

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Mattison, S.A., Blatch, G.L. & Edkins, A.L. HOP expression is regulated by p53 and RAS and characteristic of a cancer gene signature. Cell Stress and Chaperones 22, 213–223 (2017). https://doi.org/10.1007/s12192-016-0755-8

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