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Journal of Virology, March 2003, p. 3871-3877, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3871-3877.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Effects of Dipeptide Insertions between Codons 69 and 70 of Human Immunodeficiency Virus Type 1 Reverse Transcriptase on Primer Unblocking, Deoxynucleoside Triphosphate Inhibition, and DNA Chain Elongation

Peter R. Meyer,1 Johan Lennerstrand,2,3 Suzanne E. Matsuura,1 Brendan A. Larder,2,{dagger} and Walter A. Scott1*

University of Miami, School of Medicine, Miami, Florida,1 Virco, UK, Ltd., Cambridge, United Kingdom,2 Division of Clinical Virology, Huddinge University Hospital, S-141 86, Stockholm, Sweden3

Received 6 September 2002/ Accepted 12 December 2002

Finger insertion mutations of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) (T69S mutations followed by various dipeptide insertions) have a multinucleoside resistance phenotype that can be explained by decreased sensitivity to deoxynucleoside triphosphate (dNTP) inhibition of the nucleotide-dependent unblocking activity of RT. We show that RTs with SG or AG (but not SS) insertions have three- to fourfold-increased unblocking activity and that all three finger insertion mutations have threefold-decreased sensitivity to dNTP inhibition. The additional presence of M41L and T215Y mutations increased unblocking activity for all three insertions, greatly reduced the sensitivity to dNTP inhibition, and resulted in defects in in vitro DNA chain elongation. The DNA chain elongation defects were partially repaired by additional mutations at positions 210, 211, and 214. These results suggest that structural communication between the regions of RT defined by these mutations plays a role in the multinucleoside resistance phenotype.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, P.O. Box 016129, Miami, FL 33101-6129. Phone: (305) 243-6359. Fax: (305) 243-3065. E-mail: wscott{at}med.miami.edu.

{dagger} Present address: Visible Genetics, Cambridge CB4 0GA, United Kingdom.

Journal of Virology, March 2003, p. 3871-3877, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3871-3877.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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