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The 7SK small nuclear RNA inhibits the CDK9/cyclin T1 kinase to control transcription

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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Figure 1: CDK9 associates with the 7SK snRNA. a, CDK9-Flag and its associated factors (anti-Flag immunoprecipitates (IP)) were affinity purified from the nuclear extract (NE) of F1C2 cells, and analysed by SDS–polyacrylamide gel electrophoresis and silver staining.
Figure 2: SK binds to the CDK9/cyclin T1 heterodimer and inhibits P-TEFb transcriptional activity in vitro.
Figure 3: SK inhibits P-TEFb transcriptional activity in vivo.
Figure 4: Mechanism of 7SK inhibition of transcription and relief of this inhibition by stress-induced 7SK dissociation from P-TEFb. a, CDK9-Flag and its associated factors were affinity purified (anti-Flag IP) from F1C2 nuclear extract, divided into two halves, treated with RNase A or DNase I, and analysed in kinase reactions containing Pol II as a substrate.

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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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