hnRNP R negatively regulates transcription by modulating the association of P‐TEFb with 7SK and BRD4

The P‐TEFb complex promotes transcription elongation by releasing paused RNA polymerase II. P‐TEFb itself is known to be inactivated through binding to the non‐coding RNA 7SK but there is only limited information about mechanisms regulating their association. Here, we show that cells deficient in the RNA‐binding protein hnRNP R, a known 7SK interactor, exhibit increased transcription due to phosphorylation of RNA polymerase II. Intriguingly, loss of hnRNP R promotes the release of P‐TEFb from 7SK, accompanied by enhanced hnRNP A1 binding to 7SK. Additionally, we found that hnRNP R interacts with BRD4, and that hnRNP R depletion increases BRD4 binding to the P‐TEFb component CDK9. Finally, CDK9 is stabilized upon loss of hnRNP R and its association with Cyclin K is enhanced. Together, our results indicate that hnRNP R negatively regulates transcription by modulating the activity and stability of the P‐TEFb complex, exemplifying the multimodal regulation of P‐TEFb by an RNA‐binding protein.