Effects of phosphatase inhibitors on nuclease activity.
Nucleases such as DNase I, which selectively digest chromatin, are inhibited by several commonly used phosphatase inhibitors including sodium bisulfite. Two inhibitors, sodium arsenate and fructose-1,6-diphosphate, did not significantly inhibit nuclease action. Two other effective phosphatase inhibitors, p-chloromercuriphenyl sulfonate and 5,5'-dithiobis(2-nitrobenzoate), can be used during nuclei isolation and then washed out of nuclei before nuclease digestion. Using this procedure, 1mM p-chloromercuriphenyl sulfonate is as effective as 50mM bisulfite in retaining the phosphatase-sensitive mitotic phosphorylations of histones H1 and H3.
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology*; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors*; D000818:Animals; D001149:Arsenates / Q000494:pharmacology*; D001151:Arsenic / Q000494:pharmacology*; D002460:Cell Line; D002843:Chromatin / Q000648:ultrastructure; D006224:Cricetinae; D003412:Cricetulus; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology*; D005260:Female; D005635:Fructosediphosphates / Q000494:pharmacology*; D006598:Hexosediphosphates / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D009579:Nitrobenzoates / Q000494:pharmacology*; D010053:Ovary; D010663:Phenylmercury Compounds / Q000494:pharmacology*
D001149:Arsenates; D002843:Chromatin; D005635:Fructosediphosphates; D006598:Hexosediphosphates; D009579:Nitrobenzoates; D010663:Phenylmercury Compounds; D002731:4-Chloromercuribenzenesulfonate; C009277:sodium arsenate; D004228:Dithionitrobenzoic Acid; D003850:Deoxyribonuclease I; D000469:Alkaline Phosphatase; C029063:fructose-1,6-diphosphate; D001151:Arsenic