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Journal of Virology, December 2001, p. 11874-11880, Vol. 75, No. 23
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.23.11874-11880.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

RNase H Cleavage of the 5' End of the Human Immunodeficiency Virus Type 1 Genome

Hong-Qiang Gao,1,dagger Stefan G. Sarafianos,2 Edward Arnold,2 and Stephen H. Hughes1,3,*

ABL-Basic Research Program1 and HIV Drug Resistance Program,3 National Cancer Institute---Frederick, Frederick, Maryland 21702-1201, and Center for Advanced Biotechnology and Medicine and Chemistry Department, Rutgers University, Piscataway, New Jersey 08854-56382

Received 21 June 2001/Accepted 22 August 2001

The synthesis of retroviral DNA is initiated near the 5' end of the RNA. DNA synthesis is transferred from the 5' end to the 3' end of viral RNA in an RNase H-dependent step. In the case of human immunodeficiency virus type 1 (HIV-1) (and certain other retroviruses that have complex secondary structures at the ends of the viral RNA), there is the possibility that DNA synthesis can lead to a self-priming event that would block viral replication. The extent of RNase H cleavage must be sufficient to allow the strand transfer reaction to occur, but not so extensive that self-priming occurs. We have used a series of model RNA substrates, with and without a 5' cap, to investigate the rules governing RNase H cleavage at the 5' end of the HIV-1 genome. These in vitro RNase H cleavage reactions produce an RNA fragment of the size needed to block self-priming but still allow strand transfer. The cleavages seen in vitro can be understood in light of the structure of HIV-1 reverse transcriptase in a complex with an RNA/DNA substrate.

* Corresponding author. Mailing address: HIV Drug Resistance Program, National Cancer Institute-FCRDC, P.O. Box B, Building 539, Room 130A, Frederick, MD 21702-1201 Phone: (301) 846-1619. Fax: (301) 846-6966. E-mail: hughes{at}ncifcrf.gov.

dagger Present address: E-Centive, Bethesda, MD 20817.

Journal of Virology, December 2001, p. 11874-11880, Vol. 75, No. 23
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.23.11874-11880.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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