title: Basic N-terminus of yeast Nhp6A regulates the mechanism of its DNA flexibility enhancement.
abstract: HMGB (high-mobility group box) proteins are members of a class of small proteins that are ubiquitous in eukaryotic cells and nonspecifically bind to DNA, inducing large-angle DNA bends, enhancing the flexibility of DNA, and likely facilitating numerous important biological interactions. To determine the nature of this behavior for different HMGB proteins, we used atomic force microscopy to quantitatively characterize the bend angle distributions of DNA complexes with human HMGB2(Box A), yeast Nhp6A, and two chimeric mutants of these proteins. While all of the HMGB proteins bend DNA to preferred angles, Nhp6A promoted the formation of higher-order oligomer structures and induced a significantly broader distribution of angles, suggesting that the mechanism of Nhp6A is like a flexible hinge more than that of HMGB2(Box A). To determine the structural origins of this behavior, we used portions of the cationic N-terminus of Nhp6A to replace corresponding HMGB2(Box A) sequences. We found that the oligomerization and broader angle distribution correlated directly with the length of the N-terminus incorporated into the HMGB2(Box A) construct. Therefore, the basic N-terminus of Nhp6A is responsible for its ability to act as a flexible hinge and to form high-order structures.
authors.affiliation: Department of Physics, Northeastern University, Boston, MA 02115, USA.
authors.forename: Jingyun
authors.initials: J
authors.lastname: Zhang
authors.forename: Micah J
authors.initials: MJ
authors.lastname: McCauley
authors.forename: L James
authors.initials: LJ
authors.lastname: Maher
authors.forename: Mark C
authors.initials: MC
authors.lastname: Williams
authors.forename: Nathan E
authors.initials: NE
authors.lastname: Israeloff
mesh_terms: D000595:Amino Acid Sequence; D004271:DNA, Fungal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D024261:HMGB2 Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D024001:HMGN Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D024261:HMGB2 Protein; D024001:HMGN Proteins; D050505:Mutant Proteins; C086471:NHP6A protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins