Acetylation and methylation sites in histone H4 from Physarum polycephalum

Abstract

Histone H4 has been isolated and purified from plasmodia of Physarum polycephalum. The four major fragments produced by hydrolysis of H4 by acetic acid were separated and the complete amino acid sequence of two of them was determined. By analogy with calf H4, these peptides are at the C-terminus and give the sequence from residue 68 to the C-terminus (residue 102). In this 35 residue sequence there are two minor differences from calf H4: (i) residue 77 is arginine in Physarum H4 and lysine in calf H4; (ii) lysine-79 is partially methylated in Physarum. Arginine occurs at position 77 in pea H4 but the occurrence of methylated lysine at position 79 has not been reported for other species. In the N-terminal region, amino acid compositions of acetic acid, tryptic and chymotryptic peptides indicate that Physarum H4 and calf H4 have identical sequences from the N-terminus to residue 47. There may be minor differences in the region from residue 46 to residue 67. The sites of acetylation were determined by Edman degradation of acetate-labelled peptide 4–17 of Physarum H4. Acetylation was observed at positions 5, 8, 12, and 16. The only other labelled peptide was the N-terminal peptide, which is not susceptible to Edman degradation and is thus probably α-N-acetylated as in most other organisms. The results confirm the conservation of H4 sequence and place Physarum H4 in an intermediate position between lower eukaryote H4, such as yeast or Tetrahymena H4, and higher eukaryote H4, such as mammalian H4 or pea H4.