title: DNA bending versus DNA end joining activity of HMGB1 protein is modulated in vitro by acetylation.
abstract: The ability of HMGB1 protein to recognize bent DNA and to induce bending in linear duplex DNA defines HMGB1 as an architectural factor. It has already been demonstrated that the binding affinity of the protein for various bent DNA structures is enhanced upon in vivo acetylation at Lys2. Here we investigate how this modification of HMGB1 affects its ability to bend DNA. We report that the modified protein cannot bend short DNA fragments but, instead, stimulates joining of the same fragments via their ends. The same properties are exhibited in vivo by acetylated HMGB1 lacking its acidic tail. Further, in vitro acetylation of the truncated protein at Lys81 (possible upon tail removal only) restores the protein's bending ability, while the level of stimulation of DNA end joining is strongly reduced. We conclude, therefore, that the ability of HMGB1 to bend DNA or to stimulate end joining is modulated in vitro by acetylation. In an attempt to explain the properties of in vivo-acetylated HMGB1, its complexes with DNA have been analyzed by both protein-DNA cross-linking and atomic force microscopy. Unlike the parental protein, bound mainly within the internal sequences, acetylated HMGB1 binds preferentially to DNA ends. We propose that the loading of acetylated protein on DNA ends accounts for both the failure to bend DNA and the stimulation of DNA end joining.
authors.affiliation: Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria.
authors.forename: Iva
authors.initials: I
authors.lastname: Ugrinova
authors.forename: Elena
authors.initials: E
authors.lastname: Mitkova
authors.forename: Cendrine
authors.initials: C
authors.lastname: Moskalenko
authors.forename: Iliya
authors.initials: I
authors.lastname: Pashev
authors.forename: Evdokia
authors.initials: E
authors.lastname: Pasheva
mesh_terms: D000107:Acetylation; D000818:Animals; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism; D024243:HMGB1 Protein / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D014407:Tumor Cells, Cultured
chemical_list: D024243:HMGB1 Protein; D004247:DNA; D011088:DNA Ligases