Document Detail


AHR, a novel acute hypoxia-response sequence, drives reporter gene expression under hypoxia in vitro and in vivo.
MedLine Citation:
PMID:  20795945     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1) is an early immediate gene. We have previously reported that ADAMTS1 was strongly induced by hypoxia. In this study, we investigated whether ADAMTS1 promoter-driven reporter signal is detectable by acute hypoxia. We constructed the GFP (green fluorescent protein) expression vector [AHR (acute hypoxia-response sequence)-GFP] under the control of ADAMTS1 promoter and compared it with the constitutive GFP-expressing vector under the control of CMV (cytomegalovirus promoter-GFP). We transduced AHR-GFP and examined whether GFP signals can be detected under the acute hypoxia. When the human umbilical vein [HUVEC (human umbilical vein endothelial cells)] was transduced under normoxia, there were few GFP signals, while CMV-GFP showed considerable GFP signals. When HUVEC was stimulated with hypoxia, GFP signals from AHR-GFP gene were induced under hypoxic conditions. Notably, the GFP signals peaked at 3 h under hypoxia. In ischaemic hind limb model, transduced AHR-GFP showed hypoxic induction of GFP signals. In summary, we have demonstrated that the AHR system induced the reporter gene expression by acute hypoxia, and its induction is transient. This is the first report showing the unique acute hypoxia-activated gene expression system.
Authors:
Mehmet Zeynel Cilek; Satoshi Hirohata; Omer Faruk Hatipoglu; Hiroko Ogawa; Toru Miyoshi; Junko Inagaki; Takashi Ohtsuki; Hiroshi Harada; Shigeshi Kamikawa; Shozo Kusachi; Yoshifumi Ninomiya
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell biology international     Volume:  35     ISSN:  1095-8355     ISO Abbreviation:  Cell Biol. Int.     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2010-12-01     Completed Date:  2011-03-08     Revised Date:  2011-04-08    
Medline Journal Info:
Nlm Unique ID:  9307129     Medline TA:  Cell Biol Int     Country:  England    
Other Details:
Languages:  eng     Pagination:  1-8     Citation Subset:  IM    
Affiliation:
Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
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MeSH Terms
Descriptor/Qualifier:
ADAM Proteins / biosynthesis,  genetics*
Animals
Cell Hypoxia
Cells, Cultured
Endothelial Cells / metabolism
Endothelium, Vascular / metabolism
Gene Expression Regulation
Genes, Reporter
Green Fluorescent Proteins / biosynthesis,  genetics
Hindlimb
Humans
Hypoxia-Inducible Factor 1 / genetics,  metabolism
Ischemia / metabolism
Mice
Mice, Inbred C57BL
Muscle, Skeletal / blood supply,  metabolism
Promoter Regions, Genetic
RNA, Messenger / biosynthesis
Chemical
Reg. No./Substance:
0/Hypoxia-Inducible Factor 1; 0/RNA, Messenger; 147336-22-9/Green Fluorescent Proteins; EC 3.4.24.-/ADAM Proteins; EC 3.4.24.-/ADAMTS1 protein, human; EC 3.4.24.-/Adamts1 protein, mouse

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