[Effect of bases noncomplementary to the template on the effectiveness of primer interaction with DNA-polymerase alpha from the human placenta].
The comparison of the Km and Vmax values for the various primers was carried out. The primers were either completely complementary to the template or contained the non-complementary bases in different positions from the 3'-end. The number of the bases from the 3'-end to the noncomplementary nucleotide but not the primers length was supposed to determine the efficiency of the interaction of the primers containing noncomplementary bases with the enzyme. The Km values for d[(pC) (pT)7] (1.2 microM), d[(pC)3(pT)7] (2.5 microM, d[(pT)2pC(pT)7] (1.4 microM)d[(pT)4pC(pT)5(4.3 microM); d[(pT)7pC(pT)2] (11 microM) are comparable with the Km values for d(pT)7 (1.4 microM); d(pT)5 (4.2 microM) and d(pT)3 (15 mkM), respectively, but not for the decathymidilate d[(Tp)9T] (0.23 microM). The complementary interaction between the first nucleotide from the 3'-end of the primer and the template appear to play the particular role in the interaction of the enzyme with the primer. The Km values for d[(pT)10pC] and d[(pA)9pC] (with the corresponding templates) are 38 and 6 times the ones for d[(Tp)10T] and d(pA)10. However, the Km values for d[(pA)9p(rib)] (0.56 microM) which contains the deoxyribozylurea residue at the 3'-end is practically equal to the Km for d(pA)9 (0.56 microM). The Vmax values for d[(pT)10pC] and d[(pA)9pC] are 1.7 and 2.3 times the values for d[(Tp)10T] and d(pA)10, respectively. The primer affinity decreases, just as its conversion rate increases when the noncomplementary base in the primer is transferred from the 5'-to 3'-end; that results in the rate of primers elongation decrease in total.
Nevinskiĭ
G A
GA
Levina
A S
AS
Frolova
E I
EI
Podust
V N
VN
D004257:DNA Polymerase II / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D009838:Oligodeoxyribonucleotides / Q000096:biosynthesis; D010920:Placenta / Q000201:enzymology; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides; D004257:DNA Polymerase II
Vliianie nekomplementarnykh matritse osnovaniĭ na éffektivnost' vzaimodeĭstviia praĭmerov s DNK-polimerazoĭ alpha iz platsenty cheloveka.