Influence of polyethylene glycol on the ligation reaction with calf thymus DNA ligases I and II.
High concentrations of the nonspecific macromolecule polyethylene glycol 6000 (PEG 6000) enabled DNA ligases I and II from calf thymus to catalyze intermolecular blunt-end ligation of duplex DNA. Intermolecular cohesive-end ligation with these enzymes was markedly stimulated in the presence of 10-16% (w/v) PEG 6000. The effect of PEG 6000 (4-16%) on the sealing of single-stranded breaks in duplex DNA with DNA ligases I and II was not appreciably stimulatory but rather inhibitory. PEG 6000 (15%) enhanced more twofold the rate of DNA ligase II-AMP complex formation, but moderately suppressed the rate of formation of DNA ligase 1-AMP complex. Polyamines and KCl inhibited blunt-end and cohesive-end ligations with DNA ligases I and II in the presence of PEG 6000.
D000818:Animals; D002417:Cattle; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011118:Polynucleotide Ligases / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D013950:Thymus Gland / Q000201:enzymology*
D013096:Spermine; D011092:Polyethylene Glycols; D011189:Potassium Chloride; D011088:DNA Ligases; D011118:Polynucleotide Ligases; D000072481:DNA Ligase ATP