Unknown

Dataset Information

0

In vitro analysis of RNA polymerase II elongation complex dynamics.


ABSTRACT: RNA polymerase II elongation complexes (ECs) were assembled from nuclear extract on immobilized DNA templates and analyzed by quantitative mass spectrometry. Time-course experiments showed that initiation factor TFIIF can remain bound to early ECs, while levels of core elongation factors Spt4-Spt5, Paf1C, Spt6-Spn1, and Elf1 remain steady. Importantly, the dynamic phosphorylation patterns of the Rpb1 C-terminal domain (CTD) and the factors that recognize them change as a function of postinitiation time rather than distance elongated. Chemical inhibition of Kin28/Cdk7 in vitro blocks both Ser5 and Ser2 phosphorylation, affects initiation site choice, and inhibits elongation efficiency. EC components dependent on CTD phosphorylation include capping enzyme, cap-binding complex, Set2, and the polymerase-associated factor (PAF1) complex. By recapitulating many known features of in vivo elongation, this system reveals new details that clarify how EC-associated factors change at each step of transcription.

SUBMITTER: Joo YJ 

PROVIDER: S-EPMC6499329 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC4581355 | biostudies-other
| S-EPMC5467213 | biostudies-literature
| S-EPMC4294624 | biostudies-literature
| S-EPMC3378035 | biostudies-literature
| S-EPMC4521995 | biostudies-literature
| S-EPMC2754188 | biostudies-literature
| S-EPMC3169135 | biostudies-literature
| S-EPMC381665 | biostudies-other
| S-EPMC3434493 | biostudies-literature
| S-EPMC5949985 | biostudies-literature