Effect of triphosphate modifications in 2′‐deoxynucleoside 5′‐triphosphates on their specificity towards various DNA polymerases

FEBS Letters

Shirokova, Elena A¹  Jasko, Maxim V¹  Krayevsky, Alexander A¹  Martynov, Boris I²  Victorova, Lyubov S¹ 

[1] Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov Str., Moscow 117984, Russia;State Research Institute of Organic Chemistry and Technology, 23, Shosse Entuziastov, Moscow 111024, Russia

关键词: DNA polymerases;    AMV reverse transcriptase;    Terminal deoxynucleotidyl transferase;    Modified deoxynucleoside 5′-triphosphate;    dNTP;    2′-deoxynucleoside 5′-triphosphates;    AMV;    avian myeloblastosis virus;    HIV;    human immunodeficiency virus;    TDT;    terminal deoxynucleotidyl transferase;    AZTTP;    3′-azido-3′-deoxythymidine 5′-triphosphate;   

DOI  :  10.1016/S0014-5793(97)00577-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Some natural and glycon-modified dNTPs with β,γ-pyrophosphate substitution at the triphosphate residue were synthesized and studied to evaluate the effect of these modifications on substrate properties of dNTPs in DNA synthesis catalyzed by human placental DNA polymerases α and β, avian myeloblastosis virus reverse transcriptase, and calf thymus terminal deoxynucleotidyl transferase. Reverse transcriptase proved to be the enzyme least specific to such modifications; the substrate activity of β,γ-methylenediphosphonate substituted dTTP and 3′-azido-3′-deoxy-dTTP decreased in the following order: CF₂=CHF>CBr₂>CFMe≫CH₂. This order is individual for each DNA polymerase. It is interesting to mention that β,γ-CBr₂ substituted dTTP is neither a substrate nor an inhibitor of DNA polymerase β. This specificity distinguishes DNA polymerase β from other DNA polymerases studied.

【 授权许可】

Unknown