The Transcription-Dependent Dissociation of P-TEFb-HEXIM1-7SK RNA Relies upon Formation of hnRNP-7SK RNA Complexes

Abstract

The positive transcription elongation factor P-TEFb controls the elongation of transcription by RNA polymerase II. P-TEFb is inactivated upon binding to HEXIM1 or HEXIM2 proteins associated with a noncoding RNA, 7SK. In response to the inhibition of transcription, 7SK RNA, as well as HEXIM proteins, is released by an unknown mechanism and P-TEFb is activated. New partners of 7SK RNA were searched for as potential players in this feedback process. A subset of heterogeneous ribonuclear proteins, hnRNPs Q and R and hnRNPs A1 and A2, were thus identified as major 7SK RNA-associated proteins. The degree of association of 7SK RNA with these hnRNPs increased when P-TEFb-HEXIM1-7SK was dissociated following the inhibition of transcription or HEXIM1 knockdown. This finding suggested that 7SK RNA shuttles from HEXIM1-P-TEFb complexes to hnRNPs. The transcription-dependent dissociation of P-TEFb-HEXIM1-7SK complexes was attenuated when both hnRNPs A1 and A2 were knocked down by small interfering RNA. As hnRNPs are known to interact transiently with RNA while it is synthesized, hnRNPs released from nascent transcripts may trap 7SK RNA and thereby contribute to the activation of P-TEFb.

We are indebted to Anne Catherine Dock-Bregeon (Strasbourg, France), Xavier Darzacq, Gaëlle Diribarne, Annemieke Michels (Lausanne, Switzerland), and Béatrice Spiluttini for fruitful discussions and help and to Gideon Dreyfuss (Philadelphia, PA), Joëlle Marie (Gif sur Yvette, France), David Price (Iowa City, IA), William F. Rigby (Lebanon, NH), Michael Sendtner (Wuerzburg, Germany), and Chris Smith (Cambridge, United Kingdom) for reagents.

This work was supported by grants from Association pour la Recherche sur le Cancer, Agence Nationale de Recherche sur le SIDA, Agence Nationale pour la Recherche, Fondation pour la Recherche Médicale.