The acetylation of transcription factor HBP1 by p300/CBP enhances p16INK4A expression

Nucleic Acids Res. 2012 Feb;40(3):981-95. doi: 10.1093/nar/gkr818. Epub 2011 Oct 3.

Abstract

HBP1 is a sequence-specific DNA-binding transcription factor with many important biological roles. It activates or represses the expression of some specific genes during cell growth and differentiation. Previous studies have exhibited that HBP1 binds to p16(INK4A) promoter and activates p16(INK4A) expression. We found that trichostatin A (TSA), an inhibitor of HDAC (histone deacetylase), induces p16(INK4A) expression in an HBP1-dependent manner. This result was drawn from a transactivation experiment by measuring relative luciferase activities of p16(INK4A) promoter with HBP1-binding site in comparison with that of the wild-type p16(INK4A) promoter by transient cotransfection with HBP1 into HEK293T cells and 2BS cells. HBP1 acetylation after TSA treatment was confirmed by immunoprecipitation assay. Our data showed that HBP1 interacted with histone acetyltransferase p300 and CREB-binding protein (CBP) and also recruited p300/CBP to p16(INK4A) promoter. HBP1 was acetylated by p300/CBP in two regions: repression domain (K297/305/307) and P domain (K171/419). Acetylation of Repression domain was not required for HBP1 transactivation on p16(INK4A). However, luciferase assay and western blotting results indicate that acetylation of P domain, especially K419 acetylation is essential for HBP1 transactivation on p16(INK4A). As assayed by SA-beta-gal staining, the acetylation of HBP1 at K419 enhanced HBP1-induced premature senescence in 2BS cells. In addition, HDAC4 repressed HBP1-induced premature senescence through permanently deacetylating HBP1. We conclude that our data suggest that HBP1 acetylation at K419 plays an important role in HBP1-induced p16(INK4A) expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • CREB-Binding Protein / metabolism*
  • Cell Line
  • Cellular Senescence
  • Cyclin-Dependent Kinase Inhibitor p16 / genetics*
  • DNA / metabolism
  • High Mobility Group Proteins / chemistry
  • High Mobility Group Proteins / metabolism*
  • Histone Deacetylase Inhibitors / pharmacology
  • Histone Deacetylases / metabolism
  • Humans
  • Hydroxamic Acids / pharmacology
  • Lysine / metabolism
  • Promoter Regions, Genetic
  • Protein Structure, Tertiary
  • Repressor Proteins / chemistry
  • Repressor Proteins / metabolism*
  • Transcriptional Activation*
  • p300-CBP Transcription Factors / metabolism*

Substances

  • Cyclin-Dependent Kinase Inhibitor p16
  • HBP1 protein, human
  • High Mobility Group Proteins
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • Repressor Proteins
  • trichostatin A
  • DNA
  • CREB-Binding Protein
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor
  • HDAC4 protein, human
  • Histone Deacetylases
  • Lysine