HIV-1 reverse transcriptase is capable of elongating derivatives of sequence specific noncomplementary oligodeoxynucleotides.
We have carried out a comparison of KM and Vmax values for various primers in the polymerization reaction catalyzed by the HIV-1 RT. The affinity of RT for complementary d(pT)6 containing two different 5'-end pyranone derivatives was 2-3 orders of magnitude higher (KM = 3-15 nM) than that of d(pT)6 (KM = 12.6 mM). Oligodeoxynucleotides (ODNs) noncomplementary to poly(A) template were not elongated by RT. However, derivatives of d(CAGGTG) containing the 5'-terminal chromone and coumarin related groups were efficient primers showing KM (30-300 nM) and Vmax (75-93%) values comparable with that for d(pT)10 (800 nM; 100%). The [d(CAGGTG)]ddT ODN derivatives were effective inhibitors of RT. The primer function of derivatives of noncomplementary ODNs appears to be due to the additional interactions of their 5'-terminal groups with the enzyme tRNA-binding site.
Martyanov
I V
IV
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Russia.
Zakharova
O D
OD
Sottofattori
E
E
Maksakova
G A
GA
Pyshnyi
D V
DV
Mazzei
M
M
Balbi
A
A
Litvak
S
S
Tarrago-Litvak
L
L
Nevinsky
G A
GA
D001665:Binding Sites; D017931:DNA Primers / Q000378:metabolism; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011071:Poly T / Q000096:biosynthesis; D012343:RNA, Transfer / Q000378:metabolism; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D013698:Templates, Genetic
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D018894:Reverse Transcriptase Inhibitors; D011071:Poly T; D012343:RNA, Transfer; D054303:HIV Reverse Transcriptase