Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/83924
Title: Expression of N-WASP is regulated by HiF1α through the hypoxia response element in the N-WASP promoter
Authors: Salvi, Amrita
Thanabalu, Thirumaran
Keywords: N-WASP
Hypoxia
Issue Date: 2016
Source: Salvi, A., & Thanabalu, T. (2017). Expression of N-WASP is regulated by HiF1α through the hypoxia response element in the N-WASP promoter. Biochemistry and Biophysics Reports, 9, 13-21.
Series/Report no.: Biochemistry and Biophysics Reports
Abstract: Cancer cell migration and invasion involves temporal and spatial regulation of actin cytoskeleton reorganization, which is regulated by the WASP family of proteins such as N-WASP (Neural- Wiskott Aldrich Syndrome Protein). We have previously shown that expression of N-WASP was increased under hypoxic conditions. In order to characterize the regulation of N-WASP expression, we constructed an N-WASP promoter driven GFP reporter construct, N-WASPpro-GFP. Transfection of N-WASPpro-GFP construct and plasmid expressing HiF1α (Hypoxia Inducible factor 1α) enhanced the expression of GFP suggesting that increased expression of N-WASP under hypoxic conditions is mediated by HiF1α. Sequence analysis of the N-WASP promoter revealed the presence of two hypoxia response elements (HREs) characterized by the consensus sequence 5′-GCGTG-3′ at -132 bp(HRE1) and at -662 bp(HRE2) relative to transcription start site (TSS). Site-directed mutagenesis of HRE1(-132) but not HRE2(-662) abolished the HiF1α induced activation of N-WASP promoter. Similarly ChIP assay demonstrated that HiF1α bound to HRE1(-132) but not HRE2(-662) under hypoxic condition. MDA-MB-231 cells but not MDA-MB-231KD cells treated with hypoxia mimicking agent, DMOG showed enhanced gelatin degradation. Similarly MDA-MB-231KD(N-WASPpro-N-WASPR) cells expressing N-WASPR under the transcriptional regulation of WT N-WASPpro but not MDA-MB-231KD(N-WASPproHRE1-N-WASPR) cells expressing N-WASPR under the transcriptional regulation of N-WASPproHRE1 showed enhanced gelatin degradation when treated with DMOG. Thus indicating the importance of N-WASP in hypoxia induced invadopodia formation. Thus, our data demonstrates that hypoxia-induced activation of N-WASP expression is mediated by interaction of HiF1α with the HRE1(-132) and explains the role of N-WASP in hypoxia induced invadopodia formation.
URI: https://hdl.handle.net/10356/83924
http://hdl.handle.net/10220/42886
DOI: 10.1016/j.bbrep.2016.10.010
Rights: © 2016 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/).
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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