Extensive and Varied Modifications in Histone H2B of Wild-Type and Histone Deacetylase 1 Mutant Neurospora crassa

• Published in: Biochemistry (Easton) Vol. 49; no. 25; pp. 5244 - 5257
• Main Authors: Anderson, D. C, Green, George R, Smith, Kristina, Selker, Eric U
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 29-06-2010
• Subjects: Algorithms; Amino Acid Sequence; Chromatography, Liquid; Electrophoresis, Gel, Two-Dimensional; Histone Deacetylase 1 - chemistry; Histone Deacetylase 1 - genetics; Histone Deacetylase 1 - metabolism; Histones - metabolism; Methylation; Molecular Sequence Data; Neurospora crassa - enzymology; Protein Processing, Post-Translational; Spectroscopy, Fourier Transform Infrared; Tandem Mass Spectrometry
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi100391w

DNA methylation is deficient in a histone deacetylase 1 (HDA1) mutant (hda-1) strain of Neurospora crassa with inactivated histone deacetylase 1. Difference two-dimensional (2D) gels identified the primary histone deacetylase 1 target as histone H2B. Acetylation was identified by LC−MS/MS at five different lysines in wild-type H2B and at 11 lysines in hda-1 H2B, suggesting Neurospora H2B is a complex combination of different acetylated species. Individual 2D gel spots were shifted by single lysine acetylations. FTICR MS-observed methylation ladders identify an ensemble of 20−25 or more modified forms for each 2D gel spot. Twelve different lysines or arginines were methylated in H2B from the wild type or hda-1; only two were in the N-terminal tail. Arginines were modified by monomethylation, dimethylation, or deimination. H2B from wild-type and hda-1 ensembles may thus differ by acetylation at multiple sites, and by additional modifications. Combined with asymmetry-generated diversity in H2B structural states in nucleosome core particles, the extensive modifications identified here can create substantial histone-generated structural diversity in nucleosome core particles.