Different phosphoisoforms of RNA polymerase II engage the Rtt103 termination factor in a structurally analogous manner.

Abstract

The carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II) orchestrates dynamic recruitment of specific cellular machines during different stages of transcription. Signature phosphorylation patterns of Y1S2P3T4S5P6S7 heptapeptide repeats of the CTD engage specific "readers." Whereas phospho-Ser5 and phospho-Ser2 marks are ubiquitous, phospho-Thr4 is reported to only impact specific genes. Here, we identify a role for phospho-Thr4 in transcription termination at noncoding small nucleolar RNA (snoRNA) genes. Quantitative proteomics reveals an interactome of known readers as well as protein complexes that were not known to rely on Thr4 for association with Pol II. The data indicate a key role for Thr4 in engaging the machinery used for transcription elongation and termination. We focus on Rtt103, a protein that binds phospho-Ser2 and phospho-Thr4 marks and facilitates transcription termination at protein-coding genes. To elucidate how Rtt103 engages two distinct CTD modifications that are differentially enriched at noncoding genes, we relied on NMR analysis of Rtt103 in complex with phospho-Thr4- or phospho-Ser2-bearing CTD peptides. The structural data reveal that Rtt103 interacts with phospho-Thr4 in a manner analogous to its interaction with phospho-Ser2-modified CTD. The same set of hydrogen bonds involving either the oxygen on phospho-Thr4 and the hydroxyl on Ser2, or the phosphate on Ser2 and the Thr4 hydroxyl, can be formed by rotation of an arginine side chain, leaving the intermolecular interface otherwise unperturbed. This economy of design enables Rtt103 to engage Pol II at distinct sets of genes with differentially enriched CTD marks.

DOI: 10.1073/pnas.1700128114

Cite this paper

@article{Nemec2017DifferentPO, title={Different phosphoisoforms of RNA polymerase II engage the Rtt103 termination factor in a structurally analogous manner.}, author={Corey M Nemec and Fan Yang and Joshua M. Gilmore and Corinna Hintermair and Y Y Ho and Sandra Chang Tseng and Martin Heidemann and Ying Zhang and Laurence Florens and Audrey P. Gasch and Dirk Eick and Michael P Washburn and Gabriele Varani and Aseem Z Ansari}, journal={Proceedings of the National Academy of Sciences of the United States of America}, year={2017}, volume={114 20}, pages={E3944-E3953} }