In vitro acetylation of HMGB-1 and -2 proteins by CBP: the role of the acidic tail

Biochemistry. 2004 Mar 16;43(10):2935-40. doi: 10.1021/bi035615y.


Histone acetyltransferases CBP, PCAF, and Tip60 have been tested for their ability to in vitro acetylate HMGB-1 and -2 proteins and their truncated forms lacking the C-terminal tail. It was found that these proteins were substrates for CBP only. Analyses of modified proteins by electrophoresis, amino acid sequencing, and mass spectrometry showed that full-length HMGB-1 and -2 were monoacetylated at Lys2. Removal of the C terminus resulted in (i) an increased incorporation of radiolabeled acetate within the proteins to a level close to that observed with histones H3/H4 and (ii) creation of a novel target site at Lys81. Acetylated and nonmodified HMGB-1 and -2 protein lacking the acidic tail were compared relative to their binding affinity to distorted DNA and the ability to bend linear DNA. Both proteins showed similar affinities to cisplatin-damaged DNA; the acetylated protein, however, was 3-fold more effective in inducing ligase-mediated circularization of a 111-bp DNA fragment. The alterations in the acetylation pattern of HMGB-1 and -2 upon removal of the C-terminal tail are regarded as a means by which the acidic domain modulates some properties of these proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Acetyltransferases / chemistry
  • Acetyltransferases / genetics
  • Animals
  • CREB-Binding Protein
  • Cyclic AMP Response Element-Binding Protein / chemistry
  • DNA Damage
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • Genetic Vectors
  • HMGB1 Protein / chemistry*
  • HMGB1 Protein / genetics
  • HMGB2 Protein / chemistry*
  • HMGB2 Protein / genetics
  • Histone Acetyltransferases
  • Humans
  • Lysine / chemistry
  • Lysine / genetics
  • Lysine Acetyltransferase 5
  • Nuclear Proteins / chemistry*
  • Nucleic Acid Conformation
  • Peptide Fragments / chemistry*
  • Peptide Fragments / genetics
  • Protein Binding / genetics
  • Protein Structure, Tertiary / genetics
  • Rats
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Trans-Activators / chemistry*


  • Cyclic AMP Response Element-Binding Protein
  • DNA-Binding Proteins
  • HMGB1 Protein
  • HMGB2 Protein
  • Nuclear Proteins
  • Peptide Fragments
  • Recombinant Proteins
  • Trans-Activators
  • Acetyltransferases
  • CREB-Binding Protein
  • CREBBP protein, human
  • Crebbp protein, rat
  • Histone Acetyltransferases
  • KAT5 protein, human
  • Lysine Acetyltransferase 5
  • Lysine