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Originally published In Press as doi:10.1074/jbc.M102441200 on May 30, 2001

J. Biol. Chem., Vol. 276, Issue 33, 31247-31256, August 17, 2001
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Initiation of HIV-1 Reverse Transcription Is Regulated by a Primer Activation Signal*

Nancy Beerens, Fedde Groot, and Ben BerkhoutDagger

From the Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, 1100 DE Amsterdam, The Netherlands

Reverse transcription of the human immunodeficiency virus type 1 (HIV-1) RNA genome appears to be strictly regulated at the level of initiation. The primer binding site (PBS), at which the tRNA<UP><SUB>3</SUB><SUP>Lys</SUP></UP> molecule anneals and reverse transcription is initiated, is present in a highly structured region of the untranslated leader RNA. Detailed mutational analysis of the U5 leader stem identified a sequence motif in the U5 region that is critical for activation of the PBS-bound tRNA<UP><SUB><RM>3</RM></SUB><SUP><RM>Lys</RM></SUP></UP> primer. This U5 motif, termed the primer activation signal (PAS), may interact with the TPsi C arm of the tRNA<UP><SUB><RM>3</RM></SUB><SUP><RM>Lys</RM></SUP></UP> primer, similar to the additional interaction proposed for the genome of Rous sarcoma virus and its tRNATrp primer. This suggests that reverse transcription is regulated by a common mechanism in all retroviruses. In HIV-1, the PAS is masked through base pairing in the U5 leader stem. This provides a mechanism for positive and negative regulation of reverse transcription. Based on structure probing of the mutant and wild-type RNAs, an RNA secondary structure model is proposed that juxtaposes the critical PAS and PBS motifs.

* This work was supported in part by the Netherlands Foundation for Chemical Research with financial aid from the Netherlands Organization for Scientific Research (NWO-CW).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Human Retrovirology, Academic Medical Center, University of Amsterdam, P. O. Box 22700, 1100 DE Amsterdam, The Netherlands. Tel.: 31-20-5664822; Fax: 31-20-6916531, E-mail: B.Berkhout@amc.uva.nl.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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