[The effect of various substituents, joined through the 5'- and 3'-ends of the primer, initiating properties during the polymerization reaction, catalyzed by AMV-revertase].
We investigated the interaction of AMV reverse transcriptase and Klenow fragment with oligonucleotide derivatives carrying different 3'- or 5'-terminal reactive groups. It was shown that the attachment of phenazinium, ethidium, and daunomycin residues to the 5'-terminal phosphate stabilized the enzyme template primer complexes, while cholesterol and hemin residues generally decreased their stability. The increased stability in solution correlated to a certain extent with the increase in affinity of the modified primers to the enzyme template complex. Coupling of bulky R residues to the primers had a weak effect on the maximal rate of primer conversion, which is likely to be a result of the lack of strong contacts between the substituents and the enzyme, and steric obstacles hindering translocation of the primer enzyme complex. We analyzed the inhibitory effect of 23 oligonucleotide derivatives (both complementary and noncomplementary to the template) with modified 3'- and 5'-ends, and revealed several analogs inhibiting polymerization catalyzed by AMV reverse transcriptase by 70-100% at 0.1-1 microM concentrations of the reagents.
Lokhova
I A
IA
Nevinskiĭ
G A
GA
Amirkhanov
N V
NV
Godovikova
T S
TS
Ivanova
E M
EM
Kavsan
V M
VM
Koshkin
A A
AA
Levina
A S
AS
Frolova
E I
EI
Zarytova
V F
VF
D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D002384:Catalysis; D002784:Cholesterol / Q000737:chemistry; D004256:DNA Polymerase I / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry*; D003630:Daunorubicin / Q000737:chemistry; D004996:Ethidium / Q000737:chemistry; D010619:Phenazines / Q000737:chemistry; D011108:Polymers; D011166:Porphyrins / Q000737:chemistry; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*
D004277:DNA, Single-Stranded; D010619:Phenazines; D011108:Polymers; D011166:Porphyrins; D002784:Cholesterol; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; D004996:Ethidium; D003630:Daunorubicin