In vitro acetylation of HMGB-1 and -2 proteins by CBP: the role of the acidic tail.
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.
Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria.
Evdokia
E
Pasheva
Mihail
M
Sarov
Kiril
K
Bidjekov
Iva
I
Ugrinova
Bettina
B
Sarg
Herbert
H
Lindner
Iliya G
IG
Pashev
D000107:Acetylation; D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics; D000818:Animals; D050882:CREB-Binding Protein; D017362:Cyclic AMP Response Element-Binding Protein / Q000737:chemistry; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors; D024243:HMGB1 Protein / Q000737:chemistry* / Q000235:genetics; D024261:HMGB2 Protein / Q000737:chemistry* / Q000235:genetics; D051548:Histone Acetyltransferases; D006801:Humans; D008239:Lysine / Q000737:chemistry / Q000235:genetics; D000074443:Lysine Acetyltransferase 5; D009687:Nuclear Proteins / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015534:Trans-Activators / Q000737:chemistry*
D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D024243:HMGB1 Protein; D024261:HMGB2 Protein; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D015534:Trans-Activators; D000123:Acetyltransferases; D050882:CREB-Binding Protein; C494363:CREBBP protein, human; C494365:Crebbp protein, rat; D051548:Histone Acetyltransferases; C098489:KAT5 protein, human; D000074443:Lysine Acetyltransferase 5; D008239:Lysine