Selective inhibition of DNA primase activity associated with DNA polymerase alpha from calf thymus.
The photo-activatable analogs of ATP, 3'-O-(4-benzoyl) benzoic adenosine 5'-triphosphate (BzATP) and 8-azidoadenosine 5'-triphosphate (8-N3-ATP) were used to study the relationship between the polymerase activity and the closely associated primase activity of calf DNA polymerase alpha. A substantial loss of DNA primase activity occurred during pre-incubation and irradiation of DNA polymerase alpha with either BzATP or 8-N3-ATP. In contrast, polymerase activity was only slightly affected. In reactions carried out after pre-incubation with BzATP or 8-N3-ATP in the absence of UV illumination, inhibition was still observed, but it could be reversed by ATP. The specificity of the inhibition for primase activity, plus the ability of ATP to act as a antagonist of BzATP and 8-N3-ATP, suggest that effective interaction of these analogs with the multisubunit polymerase-primase complex is occurring uniquely at the active site of the DNA primase.
Bodner
J B
JB
Department of Biochemistry and Cancer Center, University of Rochester, New York 14642.
Bambara
R A
RA
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D004247:DNA / Q000096:biosynthesis; D004257:DNA Polymerase II / Q000378:metabolism*; D019915:DNA Primase; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D012316:RNA Nucleotidyltransferases / Q000037:antagonists & inhibitors* / Q000528:radiation effects; D013950:Thymus Gland / Q000201:enzymology*; D014466:Ultraviolet Rays
D001386:Azides; C033901:3'-O-(4-benzoyl)benzoyladenosine 5'-triphosphate; C017040:8-azidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II