Live-cell Analysis of Endogenous GFP-RPB1 Uncovers Rapid Turnover of Initiating and Promoter-paused RNA Polymerase II

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2018 Apr 11

PMID 29632207

Citations 106

Authors

Barbara Steurer

Roel C Janssens

Bart Geverts

Marit E Geijer

Franziska Wienholz

Arjan F Theil

Jiang Chang

Shannon Dealy

Joris Pothof

Wiggert A van Cappellen

Adriaan B Houtsmuller

Jurgen A Marteijn

Affiliations

Soon will be listed here.

Abstract

Initiation and promoter-proximal pausing are key regulatory steps of RNA Polymerase II (Pol II) transcription. To study the in vivo dynamics of endogenous Pol II during these steps, we generated fully functional GFP-RPB1 knockin cells. GFP-RPB1 photobleaching combined with computational modeling revealed four kinetically distinct Pol II fractions and showed that on average 7% of Pol II are freely diffusing, while 10% are chromatin-bound for 2.4 seconds during initiation, and 23% are promoter-paused for only 42 seconds. This unexpectedly high turnover of Pol II at promoters is most likely caused by premature termination of initiating and promoter-paused Pol II and is in sharp contrast to the 23 minutes that elongating Pol II resides on chromatin. Our live-cell-imaging approach provides insights into Pol II dynamics and suggests that the continuous release and reinitiation of promoter-bound Pol II is an important component of transcriptional regulation.

Citing Articles

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