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Nucleic Acids Research, 1991, Vol. 19, No. 3 533-537
© 1991

MOLECULAR BIOLOGY

Fidelity of reverse transcriptase of the simian immunodeficiency virus from African green monkey

Andreas Manns, Herbert König1, Michael Baier1, Reinhard Kurth1 and Frank Grosse*

1Paul-Ehrlich-lnstitut D-6070 Langen, FRG Abteilung Chemie, Max-Planck-lnstitut f{euro}r experimentelle Medizin D-3400 Göttingen

*To whom correspondence should be addressed

Received October 30, 1990. Revised December 13, 1990. Accepted December 13, 1990.

The in vitro fidelity of highly purified recombinant reverse transcriptase from simian immunodeficiency virus of African green monkeys (SIVagm) was determined. By using the {Phi}{chi}174am16 reversion assay an overall error rate of 1/19,000 was determined. This is 2.4-fold higher than the overall accuracy of purified recombinant HIV-1 reverse transcriptase, measured in parallel. The evaluation of error frequencies from nucleotide pool bias studies suggest an even higher accuracy for the SIVagm-derived reverse transcriptase. TrdGMP mismatches were formed most frequently with an error rate of 1/155,000, followed by G:dGMP (1/230,000), A:dGMP (1/315,000), G:dAMP (1/340,000), T:dCMP (1/540,000), T:dTMP (1/790,000), and A:dCMP (1/1,050,000) mispairs. Thus, according to pool bias effects and depending on the mismatch under consideration SlVagm reverse transcriptase appears to be 2 to 20-fold more accurate than the homologous enzyme from the human immunodeficiency virus type 1. This higher accuracy is not due to a co-purifying exonuclaease activity. Like the enzyme from HIV-1, the simian monkey-derived enzyme was found to be devoid of a proofreading 3' to 5' exonuclease.


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