PMID- 10939628
OWN - NLM
STAT- MEDLINE
DCOM- 20001207
LR  - 20190818
IS  - 0300-8177 (Print)
IS  - 0300-8177 (Linking)
VI  - 208
IP  - 1-2
DP  - 2000 May
TI  - Regulation of endothelin-1 gene expression in human microvascular endothelial 
      cells by hypoxia and cobalt: role of hypoxia responsive element.
PG  - 53-62
AB  - Endothelin-1 (Et-1) is a vasoconstrictor peptide that plays an important role in 
      the pathophysiology of hypertension, myocardial ischemia, and other diseases. We 
      examined the mechanism of regulation the Et-1 mRNA expression in human 
      microvascular endothelial cells (HMEC-1) in response to hypoxia and cobalt. To 
      determine whether the 5'-flanking region of Et-1 gene mediate transcriptional 
      responses to cellular hypoxia, we constructed reporter plasmids in which Et-1 
      5'-flanking sequences of Et-1 gene were fused to luciferase coding sequences. 
      Constructs, which contain native Et-1 sequence 5'-AACGTGCA-3', located between 
      -118 and -125 in the opposite orientation as the transcriptional unit, mediate 
      transcriptional response to hypoxia and cobalt. This responsiveness was inhibited 
      by genistein, a tyrosine kinase selective inhibitor. Both hypoxia and cobalt 
      induced binding of HIF-1 (hypoxia inducible-1 factor) to this Et-1 hypoxia 
      responsive element in gel shift assays. Mutation in this sequence eliminated both 
      the hypoxia-induced HIF-1 binding and luciferase expression. Using the supershift 
      assay we have shown that this hypoxia responsive element binds HIF-1alpha and 
      HIF-1beta proteins. Interestingly, genistein only slightly affected HIF-1 
      binding. These results indicate that the Et-1 gene contains HIF-1 binding hypoxia 
      responsive elements which mediate transcriptional responses to hypoxia and cobalt 
      in microvascular endothelial cells. Genistein appears to inhibit this response by 
      affecting the transcriptional activity of the HIF-1 complex, without 
      significantly affecting its DNA-binding properties.
FAU - Minchenko, A
AU  - Minchenko A
AD  - Cardeza Foundation for Hematologic Research, Department of Medicine, Jefferson 
      Medical College, Thomas Jefferson University, PA 19107, USA.
FAU - Caro, J
AU  - Caro J
LA  - eng
GR  - DK34642/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Mol Cell Biochem
JT  - Molecular and cellular biochemistry
JID - 0364456
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Endothelin-1)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (HIF1A protein, human)
RN  - 0 (Hypoxia-Inducible Factor 1)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Protein Precursors)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Transcription Factors)
RN  - 3G0H8C9362 (Cobalt)
RN  - DH2M523P0H (Genistein)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - *Cell Hypoxia
MH  - Cell Line
MH  - Cobalt/*metabolism
MH  - DNA-Binding Proteins/genetics/metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Endothelin-1/biosynthesis/*genetics/metabolism
MH  - Endothelium, Vascular/cytology/*metabolism
MH  - Enzyme Inhibitors/pharmacology
MH  - *Gene Expression Regulation
MH  - Genes, Reporter
MH  - Genistein/pharmacology
MH  - Humans
MH  - Hypoxia-Inducible Factor 1
MH  - Hypoxia-Inducible Factor 1, alpha Subunit
MH  - Luciferases/genetics/metabolism
MH  - Nuclear Proteins/genetics/metabolism
MH  - Plasmids/genetics/metabolism
MH  - Protein Precursors/metabolism
MH  - RNA, Messenger/genetics/metabolism
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Response Elements/*genetics
MH  - Transcription Factors/genetics/metabolism
EDAT- 2000/08/12 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/08/12 11:00
PHST- 2000/08/12 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/08/12 11:00 [entrez]
AID - 10.1023/a:1007042729486 [doi]
PST - ppublish
SO  - Mol Cell Biochem. 2000 May;208(1-2):53-62. doi: 10.1023/a:1007042729486.