Addition of histones to histone-depleted nuclei: effect on template activity toward DNA and RNA polymerases.
The ability of histones to block the accessibility of DNA in chromatin to DNA and RNA polymerases was measured by addition of lysine-rich or arginine-rich histones to nuclei selectively depleted of these histones. By this procedure nuclei were obtained in which all of the original lysine-rich histone in the chromatin was replaced by arginine-rich histone. Conversely in other nuclei, additional lysine-rich histone replaced some of the endogenous arginine-rich histone. Lysine-rich histone was much more effective than arginine-rich histone in blocking accessibility to DNA polymerase. Both classes of histone inhibited template activity toward RNA polymerase to a similar extent. In addition to lysine-rich histone and total arginine-rich histone, phosphorylated lysine-rich histone, two fragments of lysine-rich histone produced by cleavage with N-bromosuccinamide, and fractions IIB and IV of arginine-rich histone were added to histone-depleted nuclei. With both DNA and RNA polymerases as probes, no differences in inhibition of template activity were found when native lysine-rich histone was compared to phosphorylated lysine-rich histone. Similarly, fractions IIB and IV were indistinguishable from total arginine-rich histone. On a molar basis, the carboxyl fragment of lysine-rich histone was as effective as intact lysine-rich histone only when the amino fragment was added to it. By itself, the amino portion of lysine-rich histone was without inhibitory effect in the RNA polymerase assay and resulted in only slight inhibition of template activity toward DNA polymerase.
D001120:Arginine / Q000032:analysis; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin; D004247:DNA / Q000037:antagonists & inhibitors*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D006657:Histones / Q000032:analysis / Q000494:pharmacology*; D008239:Lysine / Q000032:analysis; D013698:Templates, Genetic
D002843:Chromatin; D006657:Histones; D004247:DNA; D001120:Arginine; D004254:DNA Nucleotidyltransferases; D012321:DNA-Directed RNA Polymerases; D008239:Lysine