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Molecular and Cellular Biology, January 2006, p. 630-642, Vol. 26, No. 2
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.2.630-642.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Regulation of Polymerase II Transcription by 7SK snRNA: Two Distinct RNA Elements Direct P-TEFb and HEXIM1 Binding

Sylvain Egloff,1 Elodie Van Herreweghe,1 and Tamás Kiss1,2*

Laboratoire de Biologie Moléculaire Eucaryote du CNRS, UMR5099 and Université Paul Sabatier, IFR109, 118 route de Narbonne, 31062 Toulouse Cedex 4, France,1 Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary2

Received 29 September 2005/ Accepted 17 October 2005

The positive transcription elongation factor b (P-TEFb), a complex of Cdk9 and cyclin T1/T2, stimulates transcription by phosphorylating RNA polymerase II. The 7SK small nuclear RNA, in cooperation with HEXIM1 protein, functions as a general polymerase II transcription regulator by sequestering P-TEFb into a large kinase-inactive 7SK/HEXIM1/P-TEFb complex. Here, determination and characterization of the functionally essential elements of human 7SK snRNA directing HEXIM1 and P-TEFb binding led to a new model for the assembly of the 7SK/HEXIM1/P-TEFb regulatory complex. We demonstrate that two structurally and functionally distinct protein binding elements located in the 5'- and 3'-terminal hairpins of 7SK support the in vivo recruitment of HEXIM1 and P-TEFb. Consistently, a minimal regulatory RNA composed of the 5' and 3' hairpins of 7SK can modulate polymerase II transcription in HeLa cells. HEXIM1 binds independently and specifically to the G24-C48/G60-C87 distal segment of the 5' hairpin of 7SK. Binding of HEXIM1 is a prerequisite for association of P-TEFb with the G302-C324 apical region of the 3' hairpin of 7SK that is highly reminiscent of the human immunodeficiency virus transactivation-responsive RNA.

* Corresponding author. Mailing address: Laboratoire de Biologie Moléculaire Eucaryote du CNRS, UMR5099 and Université Paul Sabatier, IFR109, 118 route de Narbonne, 31062 Toulouse Cedex 4, France. Phone: 33 5 61 33 59 07. Fax: 33 5 61 33 58 86. E-mail: tamas{at}ibcg.biotoul.fr.

Molecular and Cellular Biology, January 2006, p. 630-642, Vol. 26, No. 2
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.2.630-642.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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