Document Detail


Molecular identification of the gene encoding porcine tristetraprolin (TTP).
MedLine Citation:
PMID:  20721683     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
Tristetraprolin (TTP) is a CCCH tandem zinc finger protein that can bind to and destabilize certain mRNAs containing AU-rich element (ARE) binding sites. In this study, a novel porcine cDNA has been isolated by expressed sequence tag assembly and subsequently confirmed by RT-PCR analysis, and designated porcine TTP (poTTP). The open reading frame of the poTTP cDNA is 981 bp, encoding 326 amino acids. The poTTP gene is approximately 2.5 kb in size and contains a single intron. Southern blotting analysis demonstrated that it is a single copy gene. Real-time quantitative PCR analysis revealed that the poTTP gene is constitutively expressed in all detected tissues, and with the highest mRNA level in lymphoid tissues spleen and thymus. Recombinant His(6)-tagged poTTP protein and its two zinc finger mutants (C146G and H127I) were efficiently expressed and purified from Escherichia coli BL21 (DE3), respectively. In vitro, RNA-electrophoretic mobility shift assay confirmed a direct interaction between poTTP protein and porcine TNF-α (poTNF-α) mRNA ARE probe; this interaction was eliminated when using either two zinc finger mutants of poTTP. Consistently, mutations within the ARE region prevented the binding interaction between recombinant poTTP protein and poTNF-α mRNA ARE probe. These results indicate that poTTP is an ARE-binding protein that might regulate the turnover of certain mRNAs in vivo.
Authors:
Zheng-Bing Guan; Yan Shui; Jian Lu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-08-19
Journal Detail:
Title:  Molecular and cellular biochemistry     Volume:  345     ISSN:  1573-4919     ISO Abbreviation:  Mol. Cell. Biochem.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-11-05     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0364456     Medline TA:  Mol Cell Biochem     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  145-51     Citation Subset:  IM    
Affiliation:
School of Biotechnology, Jiangnan University, Wuxi, 214122, People's Republic of China. guanzhengbing@yahoo.com.cn
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