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Journal of Virology, January 2008, p. 719-727, Vol. 82, No. 2
0022-538X/08/$08.00+0     doi:10.1128/JVI.02611-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Mutations in the U5 Region Adjacent to the Primer Binding Site Affect tRNA Cleavage by Human Immunodeficiency Virus Type 1 Reverse Transcriptase In Vivo{triangledown}

Jangsuk Oh,1 Mary Jane McWilliams,1 John G. Julias,2 and Stephen H. Hughes1*

HIV Drug Resistance Program, National Cancer Institute at Frederick,1 Basic Research Program, SAIC-Frederick, Inc., Frederick, Maryland 21702-12012

Received 27 November 2006/ Accepted 23 October 2007

In retroviruses, the first nucleotide added to the tRNA primer defines the end of the U5 region in the right long terminal repeat, and the subsequent removal of this tRNA primer by RNase H exactly defines the U5 end of the linear double-stranded DNA. In most retroviruses, the entire tRNA is removed by RNase H cleavage at the RNA/DNA junction. However, the RNase H domain of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase cleaves the tRNA 1 nucleotide from the RNA/DNA junction at the U5/primer binding site (PBS) junction, which leaves an rA residue at the U5 terminus. We made sequence changes at the end of the U5 region adjacent to the PBS in HIV-1 to determine whether such changes affect the specificity of tRNA primer cleavage by RNase H. In some of the mutants, RNase H usually removed the entire tRNA, showing that the cleavage specificity was shifted by 1 nucleotide. This result suggests that the tRNA cleavage specificity of the HIV-1 RNase domain H depends on sequences in U5.

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

{triangledown} Published ahead of print on 7 November 2007.

Journal of Virology, January 2008, p. 719-727, Vol. 82, No. 2
0022-538X/08/$08.00+0     doi:10.1128/JVI.02611-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Chang, K. W., Oh, J., Alvord, W. G., Hughes, S. H. (2008). The Effects of Alternate Polypurine Tracts (PPTs) and Mutations of Sequences Adjacent to the PPT on Viral Replication and Cleavage Specificity of the Rous Sarcoma Virus Reverse Transcriptase. J. Virol. 82: 8592-8604 [Abstract] [Full Text]  


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