RGD Reference Report - Oxidoreductive modification of two cysteine residues in paired domain by Ref-1 regulates DNA-binding activity of Pax-8. - Rat Genome Database

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Oxidoreductive modification of two cysteine residues in paired domain by Ref-1 regulates DNA-binding activity of Pax-8.

Authors: Cao, X  Kambe, F  Ohmori, S  Seo, H 
Citation: Cao X, etal., Biochem Biophys Res Commun 2002 Sep 20;297(2):288-93.
RGD ID: 727430
Pubmed: PMID:12237116   (View Abstract at PubMed)

We have reported reversible oxidoreductive regulation of DNA-binding activity of Pax-8: oxidation inhibits its binding and subsequent reduction restores the binding. Here, we show that Cys-45 and Cys-57 in the paired domain of rat Pax-8, which are conserved in all Pax members, are responsible for the redox regulation of its binding. Electrophoretic mobility shift assay using deletion mutants and mutants with substitution of cysteine with serine revealed that oxidation by diamide of either Cys-45 or Cys-57 loses the DNA binding of Pax-8. An intracellular oxidoreductive enzyme redox factor-1 (Ref-1) could reduce the oxidized Cys-45 or Cys-57 and restored the binding. Furthermore, reporter gene assay showed that transcriptional activity of wild-type Pax-8 was enhanced by co-expression of Ref-1. When the mutant with double substitutions of Cys-45 and Cys-57, which was insensitive to oxidation, was transfected, the basal transactivation level was much higher than that of wild-type Pax-8, while it was not enhanced by Ref-1. These results demonstrated that oxidoreductive modification of Cys-45 and Cys-57 via Ref-1 plays a role in redox regulation of Pax-8 in living cells.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Pax8Ratpositive regulation of transcription by RNA polymerase II  IDA  RGD 
Pax8Ratsulfur compound metabolic process  IEP dithiothreitolRGD 

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Pax8RatDNA-binding transcription activator activity, RNA polymerase II-specific  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Pax8  (paired box 8)


Additional Information