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Natural selection results in conservation of HIV-1 integrase activity despite sequence variability

Reinke, Ryana; Steffen, Nicholas R.b; Robinson, W. Edward Jra,c

Basic Science

Background: Integration of the HIV genome by integrase is absolutely required for productive infection.

Objective: To determine the role of natural selection on HIV integrase biology.

Design: To study the activities of HIV integrases from a limited panel of North American clinical isolates from HIV-infected patients and to compare these proteins with integrases from two laboratory adapted reference strains (HIVIIIRF and HIVNL4−−3).

Methods: HIV was isolated and the particle-associated RNA was reverse transcribed and sequenced. Replication kinetics of molecularly cloned viruses containing each variant integrase were studied in tissue culture. The mutant integrase proteins were expressed, purified and specific activities of the enzymes were derived for both 3′ end-processing and disintegration reactions.

Results: Despite 3–5% variability in integrase at the amino acid level, viruses showed no statistically significant differences in growth kinetics compared with the reference HIVNL4−−3 virus and only minor differences were observed in 3′ end-processing and disintegration activities. All integrase proteins demonstrated similar sensitivity to an integrase inhibitor l-chicoric acid.

Conclusions: These results demonstrate that integrase genes derived from HIV-infected individuals can differ from reference sequences but these mutations do not result in loss of function, including susceptibility to an integrase inhibitor; therefore, integrase remains an attractive target for antiviral drug design, as mutability appears to be restricted by function.

From the Departments of aMicrobiology and Molecular Genetics, bInformation and Computer Science and cPathology, University of California, Irvine California, USA.

Requests for reprints to Dr W. E. Robinson Jr, Department of Pathology, University of California, Irvine, California 92967-4800, USA.

Received: 20 July 2000;

revised: 5 January 2001; accepted: 30 January 2001.

Sponsorship: WER is a recipient of the Burroughs–Wellcome Fund Clinical Scientist Award in Translational Research. This work was supported in part by the Public Health Service: 5RO1-AI-41360 (WER) and 5T32 AI-07319 (RR) and the Burroughs-Wellcome Fund: App 2609 (WER).

© 2001 Lippincott Williams & Wilkins, Inc.