Binding of a host cell nuclear protein to the stem region of human immunodeficiency virus type 1 trans-activation-responsive RNA.
Human immunodeficiency virus type 1 (HIV-1) transcription is regulated by both viral and host cell factors. Although the viral trans-activator protein, Tat, and its cis-responsive element, trans-activation-responsive (TAR) RNA, have been identified and characterized, the mechanism of HIV-1 transcriptional regulation has not been satisfactorily described. Whereas Tat is necessary to activate transcription, additional factors, derived from the host cell, are important in regulating HIV-1 transcription. To identify such host cell-specific factors, we used an RNase protection mobility shift assay and UV cross-linking to detect a 140-kDa HeLa cell nuclear protein that binds specifically to TAR RNA. By extensive mutational analysis, we determined that the binding of this protein is dependent on both the sequence and the structure of the TAR RNA stem region. Other groups have shown that the production of prematurely terminated transcripts from the HIV-1 promoter is also dependent on the sequence and structure of the TAR RNA stem. This correlation with our results suggests that the TAR RNA stem-binding protein is involved in the production of prematurely terminated transcripts from the HIV-1 promoter and in the regulation of HIV-1 gene expression.
Rounseville
M P
MP
Graduate Program in Genetics, George Washington University Medical Center, Washington, D.C. 20037.
Kumar
A
A
D001483:Base Sequence; D004252:DNA Mutational Analysis; D015967:Gene Expression Regulation, Viral; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D006860:Hydrogen Bonding; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009687:Nuclear Proteins / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D014466:Ultraviolet Rays
D009687:Nuclear Proteins; D012367:RNA, Viral