PMID- 15199099
OWN - NLM
STAT- MEDLINE
DCOM- 20050207
LR  - 20161124
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 117
IP  - Pt 15
DP  - 2004 Jul 1
TI  - Redox state regulates HIF-1alpha and its DNA binding and phosphorylation in 
      salmonid cells.
PG  - 3201-6
AB  - Rainbow trout (Oncorhynchus mykiss) hypoxia-inducible factor-1 (HIF-1) is a 
      heterodimeric transcription factor structurally similar to mammalian HIF-1. It 
      consists of HIF-1alpha and HIF-1beta subunits, of which the HIF-1alpha subunit 
      confers the hypoxia sensitivity. HIF-1alpha is rapidly degraded by a proteasome 
      under normal oxygen (21% O2) conditions, mainly as a result of prolyl 
      hydroxylation needed for protein destabilization. Although prolyl hydroxylation 
      at conserved proline residues is a major factor controlling HIF-1alpha stability, 
      the redox state of the cells may, in addition, influence the function of 
      HIF-1alpha like proteins by influencing their stability, DNA binding and 
      phosphorylation. Sensitivity of the protein to oxidation/reduction may be due to 
      cysteine residues at critical positions. The predicted amino acid sequence of 
      rainbow trout HIF-1alpha contains several unique cysteine residues, notably in 
      the DNA-binding area at position 28 and in the transactivation domain of the 
      molecule in the vicinity of the conserved proline residue at position 564 of 
      mammalian HIF-1alpha. In the present studies we have investigated if the redox 
      state influences HIF-1alpha stability, DNA binding and phosphorylation in two 
      established salmonid cell lines RTG-2 and CHSE-214. The results indicate that 
      reducing conditions, achieved using N-propylgallate (nPG) or N-acetylcysteine 
      (NAC), stabilize HIF-1alpha, facilitate its DNA binding, and increase its 
      phosphorylation even under normal oxygen conditions. On the other hand, oxidizing 
      conditions, achieved using L-buthionine sulfoximine (BSO) dampen the hypoxia 
      response. Furthermore, the hypoxia-like effect of cobalt is increased in the 
      presence of the reducing agent. On the basis of these results, we suggest that 
      redox state influences the accessibility of the conserved prolyl residues to 
      oxygen-dependent hydroxylation and the accessibility of the residues involved in 
      the phosphorylation of HIF-1alpha.
FAU - Nikinmaa, Mikko
AU  - Nikinmaa M
AD  - Department of Biology, University of Turku, 20014, Finland. miknik@utu.fi
FAU - Pursiheimo, Saijaliisa
AU  - Pursiheimo S
FAU - Soitamo, Arto J
AU  - Soitamo AJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20040615
PL  - England
TA  - J Cell Sci
JT  - Journal of cell science
JID - 0052457
RN  - 0 (Antimetabolites)
RN  - 0 (Antioxidants)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Transcription Factors)
RN  - 5072-26-4 (Buthionine Sulfoximine)
RN  - 63231-63-0 (RNA)
RN  - 8D4SNN7V92 (Propyl Gallate)
RN  - 9007-49-2 (DNA)
RN  - 9DLQ4CIU6V (Proline)
RN  - EC 3.4.25.1 (Proteasome Endopeptidase Complex)
RN  - K848JZ4886 (Cysteine)
RN  - S88TT14065 (Oxygen)
RN  - WYQ7N0BPYC (Acetylcysteine)
SB  - IM
MH  - Acetylcysteine/pharmacology
MH  - Animals
MH  - Antimetabolites/pharmacology
MH  - Antioxidants/pharmacology
MH  - Buthionine Sulfoximine/pharmacology
MH  - Cell Line
MH  - Cysteine/chemistry
MH  - DNA/*chemistry/*metabolism
MH  - Dimerization
MH  - Dose-Response Relationship, Drug
MH  - Enzyme Inhibitors/pharmacology
MH  - Hydroxylation
MH  - Hypoxia
MH  - Hypoxia-Inducible Factor 1, alpha Subunit
MH  - Immunoblotting
MH  - Immunoprecipitation
MH  - Oncorhynchus mykiss
MH  - *Oxidation-Reduction
MH  - Oxidative Stress
MH  - Oxygen/metabolism
MH  - Phosphorylation
MH  - Proline/metabolism
MH  - Propyl Gallate/pharmacology
MH  - Proteasome Endopeptidase Complex/metabolism
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - RNA/chemistry
MH  - Time Factors
MH  - Transcription Factors/*metabolism
MH  - Transcriptional Activation
EDAT- 2004/06/17 05:00
MHDA- 2005/02/08 09:00
CRDT- 2004/06/17 05:00
PHST- 2004/06/17 05:00 [pubmed]
PHST- 2005/02/08 09:00 [medline]
PHST- 2004/06/17 05:00 [entrez]
AID - jcs.01192 [pii]
AID - 10.1242/jcs.01192 [doi]
PST - ppublish
SO  - J Cell Sci. 2004 Jul 1;117(Pt 15):3201-6. doi: 10.1242/jcs.01192. Epub 2004 Jun 
      15.