Tetrahymena histone H3. Purification and two variant sequences.

The H3 histone of the protozoan Tetrahymena pyriformis was obtained as described previously (Fusauchi, Y. & Iwai, K. (1983) J. Biochem. 93, 1487-1497) and further purified by Sephadex G-50 chromatography after reduction and carboxymethylation. The purified H3 was shown to comprise two variants, 75 mol% of H3(1) and 25 mol% of H3(2). The H3 mixture was directly sequenced by Edman degradation from the N-terminal through residue 104. Sequence determination was further performed with tryptic peptides and cyanogen bromide fragments derived from the H3 mixture. Thus, the total sequences of H3(1) and H3(2) were completely determined; both consist of a total of 135 amino acid residues (the molecular weights in the unmodified form are 15,336 for H3(1) and 15,424 for H3(2), and both are partially acetylated or methylated at the same six lysine residues to similar extents. The H3(1) and H3(2) sequences differ in 14 positions from each other, and in 17 and 21 positions from those of human spleen H3 (Ohe, Y. & Iwai, K. (1981) J. Biochem. 90, 1205-1211). The implications of these results for the structure-function relationship of this histone species and also for the phylogeny of protozoa are discussed.