Interaction of polymerases with 2′-deoxyuridine-5′-triphosphate spin-labeled at the 5-position

• Published in: FEBS letters 1983-02, Vol.152 (2), p.157-162
• Main Authors: Warwick-Koochaki, P.E., Hakan, A., Bobst, A.M.
• Format: Article
• Language: English
• Subjects: 2′-deoxy-uridine-5′-triphosphate substituted at the 5-position with N-[1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl]-0; 5-trifluoromethyl-2′-deoxyuridine-5′-triphosphate; AMV; Animals; araCTP; avian myeloblastosis virus; cytosine-D-arabino-furanoside-5′-triphosphate; Deoxyuracil Nucleotides - metabolism; DNA polymerase; DNA polymerase (RNA-dependent); DNA-Directed DNA Polymerase - metabolism; DNA-Directed RNA Polymerases - metabolism; dUDP; electron spin resonance; Escherichia coli - enzymology; ESR; F3dTTP; Kinetics; Liver Neoplasms, Experimental - enzymology; Orthomyxoviridae - enzymology; pppDUGT; Reverse transcriptase inhibition; RNA polymerase; RNA-Directed DNA Polymerase - metabolism; Spin Labels; Spin-labeled nucleotide
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(83)80369-X

2′-Deoxyuridine-5′-triphosphate spin-labeled at the 5-position with N-[1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl]-O- was found to be an inhibitor of some DNA and RNA polymerases including avian myeloblastosis virus reverse transcriptase. Furthermore, the spin-labeled nucleotide was found to be incorporated internally into polydeoxythymidylic acid via reverse transcriptase to an extent of 1.0 spin-labeled base per 10 3 bases. The incorporation, monitored by electron spin resonance, is analogous to some other nucleotide inhibitors of polymerases, and the results indicate that it may be feasible to obtain sequence specific, spin-labeled DNA, enzymatically.