Document Detail


NRF-1, and AP-1 regulate the promoter of the human calpain small subunit 1 (CAPNS1) gene.
MedLine Citation:
PMID:  18234454     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Ubiquitously expressed micro- and m-calpain are cysteine proteases with broad functions in cell spreading, migration, proliferation, apoptosis, and in tumor invasion. They are heterodimers, with a distinct large 80-kDa catalytic, and a common small 28-kDa regulatory subunit (Capn4/CAPNS1). CAPNS1 is required to maintain stability and activity of both calpains. Despite its biological importance, the transcriptional regulation of this gene has not been studied, and the CAPNS1 promoter has not yet been characterized. In this study, we identified the main transcriptional start site, and cloned and characterized the ~2.0 kb upstream region of the CAPNS1 gene. Deletion analysis identified the core promoter located within region -187/+174. Site-directed mutagenesis, EMSA- and supershift analysis identified Sp1-, NRF-1-, and AP-1-binding elements within the CAPNS1 core promoter. Binding of NRF-1, Sp1 and AP-1 to the natural core promoter was confirmed by chromatin immunoprecipitation (ChIP). Site-directed mutagenesis at the NRF-1 site in HeLa and MCF7 cells substantially reduced core promoter activity by 70%, whereas mutation of the AP-1-binding and Sp1-binding site reduced promoter activity by 50% and 30%, respectively. Double mutation of the NRF-1 and the AP-1 site reduced promoter activity by 90%. In Drosophila SL2 cells, ectopic expression of NRF-1 led to a significant induction of CAPNS1 promoter activity. Furthermore, an siRNA against NRF-1 substantially reduced promoter activity in HeLa cells, which was paralleled by a significant downregulation of CAPNS1 mRNA. These results reveal that especially NRF-1, along with AP-1 and, to a minor extent, an Sp1 site, is essential for human CAPNS1 promoter activity and gene expression.
Authors:
Irfan A Asangani; Suhail A K Rasheed; J?rg H Leupold; Stefan Post; Heike Allgayer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-12-23
Journal Detail:
Title:  Gene     Volume:  410     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  2008 Feb 
Date Detail:
Created Date:  2008-02-22     Completed Date:  2008-06-02     Revised Date:  2010-05-20    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  197-206     Citation Subset:  IM    
Affiliation:
Department Experimental Surgery Mannheim, University Heidelberg, and Molecular Oncology of Solid Tumors Unit, German Cancer Research Center (DKFZ) Heidelberg, Germany.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Calpain / genetics*
DNA
DNA Primers
Humans
Molecular Sequence Data
Mutagenesis, Site-Directed
Nuclear Respiratory Factor 1 / physiology*
Promoter Regions, Genetic*
RNA Interference
Transcription Factor AP-1 / physiology*
Chemical
Reg. No./Substance:
0/DNA Primers; 0/NRF1 protein, human; 0/Nuclear Respiratory Factor 1; 0/Transcription Factor AP-1; 9007-49-2/DNA; EC 3.4.22.-/Calpain; EC 3.4.22.52/CAPNS1 protein, human

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