Open AccessAmino acid sequence of histone H1 at the ADP‐ribose‐accepting site and ADP‐ribose · histone‐H1 adduct as an inhibitor of cyclic‐AMP‐dependent phosphorylation
Author(s) -
USHIROYAMA Takahisa,
TANIGAWA Yoshinori,
TSUCHIYA Mikako,
MATSUURA Ryoji,
UEKI Minoru,
SUGIMOTO Osamu,
SHIMOYAMA Makoto
Publication year - 1985
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1985.tb09082.x
Subject(s) - histone h1 , histone h2a , histone octamer , biochemistry , histone methyltransferase , microbiology and biotechnology , adp ribosylation , histone h4 , biology , histone , phosphorylation , nad+ kinase , nucleosome , enzyme , dna
The ADP‐ribosylation site of histone H1 from calf thymus by purified hen liver nuclear ADP‐ribosyltransferase was determined and effects of the ADP‐ribose · histone‐H1 adduct on cAMP‐dependent phosphorylation of the histone H1 were investigated. ADP‐ribosylated histone H1 was prepared by incubation of histone H1, 1 mM [ adenylate ‐ 32 P]NAD and the purified ADP‐ribosyltransferase. N ‐Bromosuccinimide‐directed bisection of ADP‐ribosylated histone H1 showed that the NH 2 ‐terminal fragment ( M r = 6000) was modified and contained serine residue 38, the site of phosphorylation by cAMP‐dependent protein kinase. Digestion of the NH 2 ‐terminal fragment with cathepsin D and trypsin, and purification of this fragment, using high‐performance liquid chromatography, yielded a radiolabelled single peptide corresponding to residues 29–34 of histone H1, containing the arginine residue as the ADP‐ribosylation site. These results indicate that ADP‐ribosylation of histone H1 occurs at the arginine residue 34, sequenced at the NH 2 ‐terminal side of the phosphate‐accepting serine residue 38. Phosphorylation of histone H1 from calf thymus by cAMP‐dependent protein kinase was markedly reduced when histone H1 was ADP‐ribosylated. Kinetic studies of phosphorylation revealed that ADP‐ribosylated histone H1 was a linear competitive inhibitor of histone H1 and a linear non‐competitive inhibitor of ATP.