[RNA-dependent DNA-polymerase from avian myeloblastosis virus: effectiveness of interaction with oligothymidylate primers of various length].
Optimal conditions for the reaction of polymerization catalyzed by RNA-dependent DNA-polymerase from AMV on poly(A)- and poly(dA)-templates with d(pT)n-primers were established. Optimal concentrations of the components and pH of the reaction mixtures were found out to differ significantly. dTTP was shown to be both a nucleotide substrate and a minimal primer of the polymerization. The Km values for d(pT)2-primer (Km = 0.11 mM and 0.54 for poly(A) and poly(dA)-templates, respectively) and longer oligothymidylates were estimated. The lengthening of d(pT)n (n = 2-10) by one mononucleotide unit led to a 3-fold and 2-fold decrease of Km value for poly(A) and poly(dA), respectively. Further lengthening of the primer (n = 10-25) did not affect Km for the primers. The maximal rates of polymerization did not depend on primer length. The activation reaction (Ea = 12 kcal/mol) of polymerization on poly(A) was considerably lower than that on poly(dA) (Ea = 50 kcal/mol). In both cases a highly processive polymerization was observed. It was suggested that the synthesis had been more effective on poly(A)-template due to a more effective formation of the complex enzyme primer template.
Lokhova
I A
IA
Nevinskiĭ
G A
GA
Bulychev
N A
NA
Gorn
V V
VV
Levina
A S
AS
Rudenko
N K
NK
Kavsan
V M
VM
Lavrik
O I
OI
D001354:Avian Leukosis Virus / Q000201:enzymology*; D009189:Avian Myeloblastosis Virus / Q000201:enzymology* / Q000235:genetics; D002384:Catalysis; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011061:Poly A / Q000378:metabolism*; D011071:Poly T / Q000378:metabolism*; D011089:Polydeoxyribonucleotides / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D013698:Templates, Genetic
D011089:Polydeoxyribonucleotides; D011061:Poly A; D011071:Poly T; C015465:poly(dA); D012194:RNA-Directed DNA Polymerase
RNK-zavisimaia DNK-polimeraza iz virusa mieloblastoza ptits: éffektivnost' vzaimodeĭstviia s oligotimidilatami-praĭmerami razlichnoĭ dliny.