Rho-dependent transcription termination proceeds via three routes

Eunho Song, Heesoo Uhm, Palinda Ruvan Munasingha, Seungha Hwang, Yeon-Soo Seo, Jin Young Kang (), Changwon Kang () and Sungchul Hohng ()
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Eunho Song: Seoul National University
Heesoo Uhm: Seoul National University
Palinda Ruvan Munasingha: Korea Advanced Institute of Science and Technology
Seungha Hwang: Korea Advanced Institute of Science and Technology
Yeon-Soo Seo: Korea Advanced Institute of Science and Technology
Jin Young Kang: Korea Advanced Institute of Science and Technology
Changwon Kang: Korea Advanced Institute of Science and Technology
Sungchul Hohng: Seoul National University

Nature Communications, 2022, vol. 13, issue 1, 1-12

Abstract: Abstract Rho is a general transcription termination factor in bacteria, but many aspects of its mechanism of action are unclear. Diverse models have been proposed for the initial interaction between the RNA polymerase (RNAP) and Rho (catch-up and stand-by pre-terminational models); for the terminational release of the RNA transcript (RNA shearing, RNAP hyper-translocation or displacing, and allosteric models); and for the post-terminational outcome (whether the RNAP dissociates or remains bound to the DNA). Here, we use single-molecule fluorescence assays to study those three steps in transcription termination mediated by E. coli Rho. We find that different mechanisms previously proposed for each step co-exist, but apparently occur on various timescales and tend to lead to specific outcomes. Our results indicate that three kinetically distinct routes take place: (1) the catch-up mode leads first to RNA shearing for RNAP recycling on DNA, and (2) later to RNAP displacement for decomposition of the transcriptional complex; (3) the last termination usually follows the stand-by mode with displacing for decomposing. This three-route model would help reconcile current controversies on the mechanisms.

Date: 2022
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DOI: 10.1038/s41467-022-29321-5

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