The Tat proteins of human immunodeficiency virus types 1 (HIV-1) and 2 (HIV-2), termed Tat-1 and Tat-2, respectively, are essential for efficient viral replication. Tatproteins activate viral transcription by binding to the TAR RNA stem-loop structure at the 5' end of viral transcripts. We used an in vitro selection procedure to identify RNAs present in a large sequence pool that are able to bind to purified Tat-2 protein. The sequences of thc selected RNAs demonstrated a consensus feature: 20 of27 RNAs contained computer-predicted loop structures that were >50% U or C nucleotides. A selected RNA was characterized for its in vitro binding propertics to various Tat-2 proteins. This synthetic RNA was bound by wildtype Tat-2 protcins with an affinity that was only slightly lowcr than that of thc natural HIV-2 TAR RNA. Tat-2 required a wild-type RNA binding domain to bind to this synthetic RNA. This study indicatcs that in vitro selection techniqucs can be used to investigate TatproteinTAR RNA interactions.

Keywords:
HIV-2, Tatprotein, TARRNA, SELEX, Protcin-RNA interactions
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1997
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