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Journal of Virology, August 2002, p. 7868-7873, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7868-7873.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Analysis of Natural Variants of the Human Immunodeficiency Virus Type 1 gag-pol Frameshift Stem-Loop Structure

Amalio Telenti,* Raquel Martinez, Miguel Munoz, Gabriela Bleiber, Gilbert Greub, Dominique Sanglard, and Solange Peters

Division of Infectious Diseases and Institute of Microbiology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

Received 7 November 2001/ Accepted 23 April 2002

Human immunodeficiency virus type 1 uses ribosomal frameshifting for translation of the Gag-Pol polyprotein. Frameshift activities are thought to be tightly regulated. Analysis of gag p1 sequences from 270 plasma virions identified in 64% of the samples the occurrence of polymorphism that could lead to changes in thermodynamic stability of the stem-loop. Expression in Saccharomyces cerevisiae of p1-ß-galactosidase fusion proteins from 10 representative natural stem-loop variants and three laboratory mutant constructs (predicted the thermodynamic stability [{Delta}G°] ranging from -23.0 to -4.3 kcal/mol) identified a reduction in frameshift activity of 13 to 67% compared with constructs with the wild-type stem-loop ({Delta}G°, -23.5 kcal/mol). Viruses carrying stem-loops associated with greater than 60% reductions in frameshift activity presented profound defects in viral replication. In contrast, viruses with stem-loop structures associated with 16 to 42% reductions in frameshift efficiency displayed no significant viral replication deficit.

* Corresponding author. Mailing address: CHUV, 1011 Lausanne, Switzerland. Phone: 41 21 314 0550. Fax: 41 21 314 1008. E-mail: amalio.telenti{at}chuv.hospvd.ch.

Journal of Virology, August 2002, p. 7868-7873, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7868-7873.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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