Anatomy of the primase-alpha DNA polymerase reaction accomplished by nucleoprotein complexes harboring an extrachromosomal DNA identical with avian myeloblastosis virus core-bound DNA: influencing by carbonyldiphosphonate, mimosine and butylphenyl deoxyguanosine-5'-triphosphate.
Activities of the primase (Pr)-alpha DNA polymerase (pol) enzyme complex belonging to the naturally occurring reaction systems represented by special NP complexes harboring an extrachromosomal DNA identical with avian myeloblastosis virus (AMV) core-bound DNA (J. Riman, A. Sulová and K. Horská, Acta virol. 39, 149-159 (1995); J. Ríman and A. Sulová, Acta virol. 41, 181-192 (1997)) were studied in the absence and presence of carbonyldiphosphonate (COMDP), mimosine (MIMO), to it related ciclopirox olamine (CPX) and butylphenyl deoxyguanosine-5'-triphosphate (BuPdGTP). Reaction products radioactively labeled for RNA and DNA and synthesized with the common four ribonucleoside triphosphates (rNTPs) or rNTPs and deoxyribonucleoside triphosphates (dNTPs) in the reaction medium, were analyzed by polyacrylamide gel electrophoresis (PAGE) at denaturing conditions. It was shown that COMDP strongly activates the Pr and uncouples its activity from that of alpha DNA pol with accumulation of initiator (i) RNAs of the basic length. This phenomenon is not affected by BuPdGTP. MIMO, in contrast, stimulates both pol activities of this enzyme complex and preserves their mutual coupling. The effects of COMDP, MIMO and CPX seem to be modulated by concentration of the ambient dNTPs. Addition of dNTPs to rNTPs makes the effects of COMDP and MIMO mutually exclusive, suggesting that both these agents, though chemically quite different, are competing for one active site responsible for coupling these both pol activities into the one Pr-alpha DNA pol reaction.
Ríman
J
J
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
D009189:Avian Myeloblastosis Virus / Q000235:genetics* / Q000502:physiology; D001665:Binding Sites; D004256:DNA Polymerase I / Q000378:metabolism*; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors; D019915:DNA Primase / Q000378:metabolism*; D004279:DNA, Viral / Q000096:biosynthesis* / Q000187:drug effects; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology; D004164:Diphosphonates / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008898:Mimosine / Q000494:pharmacology; D009687:Nuclear Proteins / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D014758:Viral Core Proteins / Q000378:metabolism*; D014779:Virus Replication
D004279:DNA, Viral; D003848:Deoxyguanine Nucleotides; D004164:Diphosphonates; D004791:Enzyme Inhibitors; D009687:Nuclear Proteins; D011506:Proteins; D014758:Viral Core Proteins; C062324:carbonyldiphosphonate; D008898:Mimosine; C040364:N(2)-(4-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate; D019915:DNA Primase; C116932:DNA polymerase alpha-primase; D004256:DNA Polymerase I; D004257:DNA Polymerase II