Localization of the sites along nucleosome DNA which interact with NH2-terminal histone regions

Abstract

Trypsin digestion of HeLa nucleosomes produces the same series of discrete histone breakdown products observed previously by others during digestion of chromatin; thus, trypsin excises the NH2-terminal ends of the histones from the chromatin core particle. The resulting nucleoprotein complex sediments at 9 S, has an increased molecular ellipticity at 280 nm, and has DNase I-susceptible sites at 10 nucleotide intervals. Nucleosomes containing a 32P label at the 5'-DNA termini were digested sequentially with trypsin and DNase I. Following trypsin digestion, the segments of nucleosome DNA 20 to 35 and 60 to 80 nucleotides from the 5' end became more susceptible to DNase I, suggesting that these segments interact with the trypsin-sensitive regions of the histones.