The histone acetyltransferase activity of human GCN5 and PCAF is stabilized by coenzymes

J Biol Chem. 1997 Oct 24;272(43):27253-8. doi: 10.1074/jbc.272.43.27253.

Abstract

Here we report that PCAF and human GCN5, two related type A histone acetyltransferases, are unstable enzymes that under the commonly used assay conditions are rapidly and irreversibly inactivated. In addition, we report that free histone H1, although not acetylated in vivo, is a preferred and convenient in vitro substrate for the study of PCAF, human GCN5, and possibly other type A histone acetyltransferases. Using either histone H1 or histone H3 as substrates, we find that preincubation with either acetyl-CoA or CoA stabilizes the acetyltransferase activities of PCAF, human GCN5 and an enzymatically active PCAF deletion mutant containing the C-terminal half of the protein. The stabilization requires the continuous presence of coenzyme, suggesting that the acetyltransferase-coenzyme complexes are stable, while the isolated apoenzymes are not. Human GCN5 and the N-terminal deletion mutant of PCAF are stabilized equally well by preincubation with either CoA or acetyl-CoA, while intact PCAF is better stabilized by acetyl-CoA than by CoA. Intact PCAF, but not the N-terminal truncation mutant or human GCN5, is autoacetylated. These findings raise the possibility that the intracellular concentrations of the coenzymes affect the stability and therefore the nuclear activity of these acetyltransferases.

MeSH terms

  • Acetyl Coenzyme A / metabolism*
  • Acetyl-CoA C-Acyltransferase / metabolism*
  • Acetylation
  • Acetyltransferases / metabolism*
  • Cell Cycle Proteins
  • Coenzymes / metabolism*
  • Enzyme Stability
  • Histone Acetyltransferases
  • Histones / metabolism*
  • Humans
  • Kinetics
  • Saccharomyces cerevisiae Proteins*
  • Substrate Specificity
  • Trans-Activators / metabolism*
  • Transcription Factors
  • p300-CBP Transcription Factors

Substances

  • Cell Cycle Proteins
  • Coenzymes
  • Histones
  • Saccharomyces cerevisiae Proteins
  • Trans-Activators
  • Transcription Factors
  • Acetyl Coenzyme A
  • Acetyltransferases
  • Acetyl-CoA C-Acyltransferase
  • Histone Acetyltransferases
  • KAT2A protein, human
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor