Oxidative stress induces Ser 2 dephosphorylation of the RNA polymerase II CTD and premature transcription termination

ABSTRACT

The C-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II) consists of YSPTSPS heptapeptide repeats, and the phosphorylation status of the repeats controls multiple transcriptional steps and co-transcriptional events. However, how CTD phosphorylation status responds to distinct environmental stresses is not fully understood. In this study, we found that a drastic reduction in phosphorylation of a subset of Ser2 residues occurs rapidly but transiently following exposure to H₂O₂. ChIP analysis indicated that Ser2-P, and to a lesser extent Tyr1-P was reduced only at the gene 3’ end. Significantly, the levels of polyadenylation factor CstF77, as well as Pol II, were also reduced. However, no increase in uncleaved or readthrough RNA products was observed, suggesting transcribing Pol II prematurely terminates at the gene end in response to H₂O₂. Further analysis found that the reduction of Ser2-P is, at least in part, regulated by CK2 but independent of FCP1 and other known Ser2 phosphatases. Finally, the H₂O₂ treatment also affected snRNA 3’ processing although surprisingly the U2 processing was not impaired. Together, our data suggest that H₂O₂ exposure creates a unique CTD phosphorylation state that rapidly alters transcription to deal with acute oxidative stress, perhaps creating a novel “emergency brake” mechanism to transiently dampen gene expression.

KEWORDS:

• RNA polymerase II
• C-terminal domain
• Oxidative stress

Acknowledgments

We thank Jing-Ping Hsin for providing experimental materials and for helpful discussion. We also thank members of the Manley laboratory for discussion. This work was supported by NIH grant R35 GM118136 (J.L.M.). L.L. was supported by the National Science Foundation Graduate Research Fellowship under Grant No. DGE – 1644869.

Disclosure statement

No potential conflict of interest was reported by the authors.

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Funding

This work was supported by the [R35 GM118136]; [DGE – 1644869].