Nucleic Acids Research, 2000, Vol. 28, No. 23 4783-4789
© 2000 Oxford University Press
Essential regions of the tRNA primer required for HIV-1 infectivity
Department of Microbiology, University of Alabama at Birmingham, 701 South 19th Street, LHRB 619, Birmingham, AL 35294, USA
Human immunodeficiency virus (HIV), like all retroviruses, requires a cellular tRNA as a primer for initiation of reverse transcription. In a previous study, we demonstrated that an HIV-1 with a primer binding site complementary to yeast tRNAPhe (psHIV-Phe) was not infectious unless yeast tRNAPhe was supplied in trans. This unique in vivo complementation system has now been used to define the elements of the tRNA required for HIV-1 replication. Mutant tRNAPhe with deletions in T
C stem–loop, anticodon stem–loop or D stem–loop of the tRNA were generated and assessed for the capacity to rescue psHIV-Phe. Mutant tRNAPhe with disrupted TC stem–loop did not rescue psHIV-Phe. In contrast, a mutant tRNAPhe without the D stem–loop was fully functional for the rescue. The tRNA anticodon stem–loop region was found to be important for efficient complementation. The results of our studies demonstrate for the first time the importance of specific structural and sequence elements of the tRNA primer for HIV-1 reverse transcription and define new targets for interruption of HIV-1 replication.
* To whom correspondence should be addressed. Tel: +1 205 9345705; Fax: +1 205 9341580; Email: casey_morrow{at}microbio.uab.edu
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