Inhibitory effects of various 3'-deoxyribonucleotides on DNA polymerase alpha 2-primase from developing cherry salmon (Oncorhynchus masou) testes.
DNA polymerase alpha 2-primase has been purified 2750 fold from developing cherry salmon (Oncorhynchus masou) testes by the following purification steps: fractional extraction, phosphocellulose (1st), ammonium sulfate fractionation, DEAE-cellulose, phosphocellulose (2nd), hydroxylapatite and single-stranded DNA-cellulose column chromatographies. Final preparation of this enzyme has a specific activity of 107,000 units/mg protein (activated salmon sperm DNA as template-primer). DNA primase activity (rGTP dependent incorporation of labelled dGMP into poly (dC) or rNTP dependent incorporation of dNMP into M13 single-stranded DNA) was tightly associated with DNA polymerase alpha activity during all stage of this purification process. Inhibition of DNA primase activity by six kinds of 3'-deoxyribonucleotides was studied by using rNTP dependent DNA synthesis on M13 DNA as template. The inhibition constants (Ki) were larger than those of DNA-dependent RNA polymerases I and II. However, Ki/Km values were very close.
D000818:Animals; D019915:DNA Primase; D003854:Deoxyribonucleotides / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D012316:RNA Nucleotidyltransferases / Q000037:antagonists & inhibitors*; D012265:Ribonucleotides / Q000494:pharmacology*; D012474:Salmon; D013329:Structure-Activity Relationship; D013737:Testis / Q000201:enzymology*
D003854:Deoxyribonucleotides; D012265:Ribonucleotides; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases