Up-regulation of 94-kDa glucose-regulated protein by hypoxia-inducible factor-1 in human endothelial cells in response to hypoxia.
Hypoxic environment in solid tumor is known to favor cell survival and to initiate the formation of new capillaries. In this work, we identified by 2D gel analysis 94-kDa glucose-regulated protein (GRP94) as being upregulated in human endothelial cells in response to hypoxia. Three putative hypoxia responsive elements (HRE) were found in the GRP94 promoter. Competition experiments of HIF-1 DNA binding using specific probes containing each HRE sequence of the GRP94 promoter clearly evidenced that HIF-1 binds these sequences with high affinity. The human GRP94 promoter was then cloned upstream of the luciferase gene and showed enhanced activity in hypoxic conditions. Mutation of two of the three HREs present in this promoter completely inhibited the hypoxia-induced increase in luciferase activity.
Paris
Sébastien
S
Laboratory of Biochemistry and Cellular Biology, University of Namur, 61 Rue de Bruxelles, 5000 Namur, Belgium.
Denis
Hélène
H
Delaive
Edouard
E
Dieu
Marc
M
Dumont
Valéry
V
Ninane
Noëlle
N
Raes
Martine
M
Michiels
Carine
C
D001483:Base Sequence; D002457:Cell Extracts / Q000737:chemistry; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D042783:Endothelial Cells / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008565:Membrane Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism*; D015854:Up-Regulation
D002457:Cell Extracts; D004268:DNA-Binding Proteins; C497442:HIF1A protein, human; D018840:HSP70 Heat-Shock Proteins; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008565:Membrane Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C018551:glucose-regulated proteins; D008156:Luciferases