Correlating Transcription Initiation and Conformational Changes by a Single-Subunit RNA Polymerase with Near Base-Pair Resolution

Molecular Cell
Hye Ran KohTaekjip Ha

Abstract

We provide a comprehensive analysis of transcription in real time by T7 RNA Polymerase (RNAP) using single-molecule fluorescence resonance energy transfer by monitoring the entire life history of transcription initiation, including stepwise RNA synthesis with near base-pair resolution, abortive cycling, and transition into elongation. Kinetically branching pathways were observed for abortive initiation with an RNAP either recycling on the same promoter or exchanging with another RNAP from solution. We detected fast and slow populations of RNAP in their transition into elongation, consistent with the efficient and delayed promoter release, respectively, observed in ensemble studies. Real-time monitoring of abortive cycling using three-probe analysis showed that the initiation events are stochastically branched into productive and failed transcription. The abortive products are generated primarily from initiation events that fail to progress to elongation, and a majority of the productive events transit to elongation without making abortive products.

Citations

May 6, 2019·Biophysics Reviews·Matthew A B Baker
Aug 29, 2020·Nature Communications·Byeong-Kwon SohnHajin Kim
Aug 30, 2020·Nature Communications·Zhaowei LiuMichael A Nash
Sep 23, 2020·RNA·Luiz F M PassalacquaAndrej Lupták
Jul 23, 2021·Proceedings of the National Academy of Sciences of the United States of America·Dylan M PlaskonM Thomas Record
Apr 6, 2019·Biochemistry·Kate L HendersonM Thomas Record
Aug 18, 2021·The Journal of Biological Chemistry·Chun-Ying Lee, Sua Myong
Oct 30, 2019·Chemical Reviews·Sonisilpa MohapatraTaekjip Ha

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