Document Detail


Identification of a short form of ubiquitin-specific protease 3 that is a repressor of rat glutathione S-transferase gene expression.
MedLine Citation:
PMID:  16279867     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The transcription rate and protein expression from both GSTA2 (glutathione S-transferase A2) and albumin genes decrease in rat liver after IL-6 (interleukin 6) plus DEX (dexamethasone) treatment of primary hepatocytes or after LPS (lipopolysaccharide)-induced acute-phase response in animals. The down-regulation is associated with the induced expression of a nuclear protein (termed IL6DEX-NP for IL-6/DEX-induced nuclear protein) that binds to a specific site on the promoter of GSTA2, leading to a decrease in transcriptional activity. IL6DEX-NP is not similar to other transcription factors, and, for identification, we functionally cloned it from a rat liver library using a yeast one-hybrid screen based on DNA-binding activity. The cloned sequence was a truncated form of USP3 (ubiquitin-specific protease 3) and the truncated USP3 protein in a yeast extract bound to DNA containing the IL6DEX-NP recognition sequence. Using 5'- and 3'-RACE (rapid amplification of cDNA ends), the complete sequence of USP3 was found in liver from LPS-treated rats. However, using Western blot analysis, only truncated forms of USP3 could be identified in nuclear extracts from LPS-treated rat livers. A GSTA2 promoter-reporter gene plasmid and USP3-expressing plasmids were transfected into rat hepatoma cells. Expression of the short form of USP3, but not the full-length protein, abolished expression from the reporter gene. Chromatin immunoprecipitation localized USP3 to the GSTA2 promoter in rat hepatocytes in vivo. We believe that the short form of USP3 is IL6DEX-NP and that it may play an important role in the negative regulation of proteins during the acute-phase response.
Authors:
Richard Whalen; Xiangdang Liu; Thomas D Boyer
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  The Biochemical journal     Volume:  394     ISSN:  1470-8728     ISO Abbreviation:  Biochem. J.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-02-07     Completed Date:  2006-04-13     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  519-26     Citation Subset:  IM    
Affiliation:
The University of Arizona Liver Research Institute, College of Medicine, University of Arizona, Tucson, AZ 85724-5136, USA. rwhalen@u.arizona.edu
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Cell Line, Tumor
Chromatin Immunoprecipitation
Cloning, Molecular
DNA / genetics,  metabolism
Endopeptidases / chemistry*,  genetics,  metabolism*
Gene Expression Regulation, Enzymologic*
Gene Library
Glutathione Transferase / genetics*
Liver / metabolism
Molecular Sequence Data
Molecular Weight
Protein Binding
Protein Isoforms / chemistry,  genetics,  metabolism
Rats
Repressor Proteins / chemistry,  genetics,  metabolism*
Transfection
Grant Support
ID/Acronym/Agency:
GMO31555/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Protein Isoforms; 0/Repressor Proteins; 9007-49-2/DNA; EC 2.5.1.18/Glutathione Transferase; EC 3.4.-/Endopeptidases; EC 3.4.99.-/ubiquitin-specific protease
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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