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Science 19 February 1988:
Vol. 239. no. 4842, pp. 910 - 913
DOI: 10.1126/science.3277284
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Articles

Science, Vol 239, Issue 4842, 910-913
Copyright © 1988 by American Association for the Advancement of Science

articles

Site-directed mutagenesis of two trans-regulatory genes (tat-III,trs) of HIV-1

MR Sadaie, T Benter, and F Wong-Staal

Laboratory of Tumor Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.

Point mutations were introduced into the overlapping trans-regulatory genes (tat-III and trs) of human immunodeficiency virus type 1 (HIV-1), and the mutants were evaluated for virus expression. The results showed that tat-III has a positive transacting role and is required for transcriptional activation. A chain terminating mutation early in the trs gene resulted in an increase in transcription of viral messenger RNA as measured by nuclear transcription experiments, but only one major species of viral messenger RNA (1.8 kilobases) was detected, and little or no viral structural proteins were made. Thus, the trs gene product is essential for expression of virus structural proteins but, at the same time, may have a negative trans-regulatory role in transcription. Cotransfection of the point mutant proviruses defective in tat or trs with each other or with a complementary DNA clone containing tat and trs sequences restored the normal transcription pattern and subsequent virus production.


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