PMID- 9242677
OWN - NLM
STAT- MEDLINE
DCOM- 19970905
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 272
IP  - 32
DP  - 1997 Aug 8
TI  - Oxygen-regulated transferrin expression is mediated by hypoxia-inducible 
      factor-1.
PG  - 20055-62
AB  - Transferrin (Tf) is a liver-derived iron transport protein whose plasma 
      concentration increases following exposure to hypoxia. Here, we present a cell 
      culture model capable of expressing Tf mRNA in an oxygen-dependent manner. A 
      4-kilobase pair Tf promoter/enhancer fragment as well as the 300-base pair 
      liver-specific Tf enhancer alone conveyed hypoxia responsiveness to a 
      heterologous reporter gene construct in hepatoma but not HeLa cells. Within this 
      enhancer, a 32-base pair hypoxia-responsive element was identified, which 
      contained two hypoxia-inducible factor-1 (HIF-1) binding sites (HBSs). Mutation 
      analysis showed that both HBSs function as oxygen-regulated enhancers in 
      Tf-expressing as well as in non-Tf-expressing cell lines. Mutation of both HBSs 
      was necessary to completely abolish hypoxic reporter gene activation. Transient 
      co-expression of the two HIF-1 subunits HIF-1alpha and aryl hydrocarbon receptor 
      nuclear translocator (ARNT)/HIF-1beta resulted in enhanced reporter gene 
      expression even under normoxic conditions. Overexpression of a dominant-negative 
      ARNT/HIF-1beta mutant reduced hypoxic activation. DNA binding studies using 
      nuclear extracts from the mouse hepatoma cell line Hepa1 and the 
      ARNT/HIF-1beta-deficient subline Hepa1C4, as well as antibodies raised against 
      HIF-1alpha and ARNT/HIF-1beta confirmed that HIF-1 binds the Tf HBSs. Mutation 
      analysis and competition experiments suggested that the 5' HBS was more efficient 
      in binding HIF-1 than the 3' HBS. Finally, hypoxic induction of endogenous Tf 
      mRNA was abrogated in Hepa1C4 cells, confirming that HIF-1 confers oxygen 
      regulation of Tf gene expression by binding to the two HBSs present in the Tf 
      enhancer.
FAU - Rolfs, A
AU  - Rolfs A
AD  - Institute of Physiology, University of Zurich-Irchel, CH-8057 Zurich, 
      Switzerland.
FAU - Kvietikova, I
AU  - Kvietikova I
FAU - Gassmann, M
AU  - Gassmann M
FAU - Wenger, R H
AU  - Wenger RH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (ARNT protein, human)
RN  - 0 (Arnt protein, mouse)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (HIF1A protein, human)
RN  - 0 (Hif1a protein, mouse)
RN  - 0 (Hypoxia-Inducible Factor 1)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Nuclear Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Aryl Hydrocarbon)
RN  - 0 (Transcription Factors)
RN  - 0 (Transferrin)
RN  - 138391-32-9 (Aryl Hydrocarbon Receptor Nuclear Translocator)
RN  - 9007-49-2 (DNA)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - Animals
MH  - Aryl Hydrocarbon Receptor Nuclear Translocator
MH  - Binding Sites
MH  - Carcinoma, Hepatocellular/metabolism
MH  - DNA/metabolism
MH  - DNA-Binding Proteins/*metabolism
MH  - *Helix-Loop-Helix Motifs
MH  - Humans
MH  - Hypoxia-Inducible Factor 1
MH  - Hypoxia-Inducible Factor 1, alpha Subunit
MH  - Liver Neoplasms/metabolism
MH  - Mice
MH  - Nuclear Proteins/*metabolism
MH  - Oxygen/*metabolism
MH  - RNA, Messenger/metabolism
MH  - Receptors, Aryl Hydrocarbon/metabolism
MH  - Repetitive Sequences, Nucleic Acid
MH  - Transcription Factors/*metabolism
MH  - Transferrin/*genetics/metabolism
MH  - Tumor Cells, Cultured
EDAT- 1997/08/08 00:00
MHDA- 1997/08/08 00:01
CRDT- 1997/08/08 00:00
PHST- 1997/08/08 00:00 [pubmed]
PHST- 1997/08/08 00:01 [medline]
PHST- 1997/08/08 00:00 [entrez]
AID - S0021-9258(18)38889-6 [pii]
AID - 10.1074/jbc.272.32.20055 [doi]
PST - ppublish
SO  - J Biol Chem. 1997 Aug 8;272(32):20055-62. doi: 10.1074/jbc.272.32.20055.