Abstract
The human positive transcription elongation factor P-TEFb, consisting of a CDK9/cyclin T1 heterodimer, functions as both a general and an HIV-1 Tat-specific transcription factor1,2. P-TEFb activates transcription by phosphorylating RNA polymerase (Pol) II, leading to the formation of processive elongation complexes. As a Tat cofactor, P-TEFb stimulates HIV-1 transcription by interacting with Tat and the transactivating responsive (TAR) RNA structure located at the 5′ end of the nascent viral transcript3. Here we identified 7SK, an abundant and evolutionarily conserved small nuclear RNA (snRNA) of unknown function4,5, as a specific P-TEFb-associated factor. 7SK inhibits general and HIV-1 Tat-specific transcriptional activities of P-TEFb in vivo and in vitro by inhibiting the kinase activity of CDK9 and preventing recruitment of P-TEFb to the HIV-1 promoter. 7SK is efficiently dissociated from P-TEFb by treatment of cells with ultraviolet irradiation and actinomycin D. As these two agents have been shown to significantly enhance HIV-1 transcription and phosphorylation of Pol II (refs 6,7,8), our data provide a mechanistic explanation for their stimulatory effects. The 7SK/P-TEFb interaction may serve as a principal control point for the induction of cellular and HIV-1 viral gene expression during stress-related responses. Our studies demonstrate the involvement of an snRNA in controlling the activity of a Cdk–cyclin kinase.
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References
Jones, K. A. Taking a new TAK on Tat transactivation. Genes Dev. 11, 2593–2599 (1997).
Price, D. H. P-TEFb, a cyclin-dependent kinase controlling elongation by RNA polymerase II. Mol. Cell. Biol. 20, 2629–2634 (2000).
Wei, P., Garber, M. E., Fang, S. M., Fischer, W. H. & Jones, K. A. A novel CDK9-associated C-type cyclin interacts directly with HIV-1 Tat and mediates its high-affinity, loop-specific binding to TAR RNA. Cell 92, 451–462 (1998).
Wassarman, D. A. & Steitz, J. A. Structural analyses of the 7SK ribonucleoprotein (RNP), the most abundant human small RNP of unknown function. Mol. Cell. Biol. 11, 3432–3445 (1991).
Zieve, G. & Penman, S. Small RNA species of the HeLa cell: metabolism and subcellular localization. Cell 8, 19–31 (1976).
Cassé, C., Giannoni, F., Nguyen, V. T., Dubois, M. F. & Bensaude, O. The transcriptional inhibitors, actinomycin D and α-amanitin, activate the HIV-1 promoter and favor phosphorylation of the RNA polymerase II C-terminal domain. J. Biol. Chem. 274, 16097–16106 (1999).
Kumar, S. et al. Activation of the HIV-1 long terminal repeat by cytokines and environmental stress requires an active CSBP/p38 MAP kinase. J. Biol. Chem. 271, 30864–30869 (1996).
Valerie, K. et al. Activation of human immunodeficiency virus type 1 by DNA damage in human cells. Nature 333, 78–81 (1988).
Zhou, Q., Chen, D., Pierstorff, E. & Luo, K. Transcription elongation factor P-TEFb mediates Tat activation of HIV-1 transcription at multiple stages. EMBO J. 17, 3681–3691 (1998).
Black, D. L., Chabot, B. & Steitz, J. A. U2 as well as U1 small nuclear ribonucleoproteins are involved in premessenger RNA splicing. Cell 42, 737–750 (1985).
Zieve, G., Benecke, B. J. & Penman, S. Synthesis of two classes of small RNA species in vivo and in vitro. Biochemistry 16, 4520–4525 (1977).
Murphy, S. et al. DNA sequences complementary to human 7 SK RNA show structural similarities to the short mobile elements of the mammalian genome. J. Mol. Biol. 177, 575–590 (1984).
Zhou, Q. & Sharp, P. A. Novel mechanism and factor for regulation by HIV-1 Tat. EMBO J. 14, 321–328 (1995).
Luo, Y., Kurz, J., MacAfee, N. & Krause, M. O. C-myc deregulation during transformation induction: involvement of 7SK RNA. J. Cell. Biochem. 64, 313–327 (1997).
Schnapp, G., Rodi, H. P., Rettig, W. J., Schnapp, A. & Damm, K. One-step affinity purification protocol for human telomerase. Nucleic Acids Res. 26, 3311–3313 (1998).
Ping, Y. H. & Rana, T. M. Tat-associated kinase (P-TEFb): a component of transcription preinitiation and elongation complexes. J. Biol. Chem. 274, 7399–7404 (1999).
Zhou, M. et al. Tat modifies the activity of CDK9 to phosphorylate serine 5 of the RNA polymerase II carboxyl-terminal domain during human immunodeficiency virus type 1 transcription. Mol. Cell. Biol. 20, 5077–5086 (2000).
Fong, Y. W. & Zhou, Q. Relief of two built-in autoinhibitory mechanisms in P-TEFb is required for assembly of a multicomponent transcription elongation complex at the human immunodeficiency virus type 1 promoter. Mol. Cell. Biol. 20, 5897–5907 (2000).
Boyd, K. E., Wells, J., Gutman, J., Bartley, S. M. & Farnham, P. J. c-Myc target gene specificity is determined by a post-DNA binding mechanism. Proc. Natl Acad. Sci. USA 95, 13887–13892 (1998).
Kretz-Remy, C. & Arrigo, A. P. The kinetics of HIV-1 long terminal repeat transcriptional activation resemble those of hsp70 promoter in heat-shock treated HeLa cells. FEBS Lett. 353, 339–344 (1994).
Vlach, J. et al. Induction of Sp1 phosphorylation and NF-κB-independent HIV promoter domain activity in T lymphocytes stimulated by okadaic acid. Virology 208, 753–761 (1995).
Weinberg, R. A. & Penman, S. Small molecular weight monodisperse nuclear RNA. J. Mol. Biol. 38, 289–304 (1968).
Lis, J. T., Mason, P., Peng, J., Price, D. H. & Werner, J. P-TEFb kinase recruitment and function at heat shock loci. Genes Dev. 14, 792–803 (2000).
Peter, M. The regulation of cyclin-dependent kinase inhibitors (CKIs). Prog. Cell Cycle Res. 3, 99–108 (1997).
Acknowledgements
We thank L. Jin for technical assistance, I. von Reis for reagent, O. Bensaude for communicating his work before publication, and Y. Fong and S. Stroschein for critical comments on the manuscript. Supported by grants from the National Institute of Health, the American Cancer Society, the Hellman Faculty Fund and the France-Berkeley Fund to Q.Z.
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Yang, Z., Zhu, Q., Luo, K. et al. The 7SK small nuclear RNA inhibits the CDK9/cyclin T1 kinase to control transcription. Nature 414, 317–322 (2001). https://doi.org/10.1038/35104575
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DOI: https://doi.org/10.1038/35104575
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