Helicase promotes replication re-initiation from an RNA transcript

Bo Sun (), Anupam Singh, Shemaila Sultana, James T. Inman, Smita S. Patel and Michelle D. Wang ()
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Bo Sun: Cornell University
Anupam Singh: Robert Wood Johnson Medical School, Rutgers University
Shemaila Sultana: Robert Wood Johnson Medical School, Rutgers University
James T. Inman: Cornell University
Smita S. Patel: Robert Wood Johnson Medical School, Rutgers University
Michelle D. Wang: Cornell University

Nature Communications, 2018, vol. 9, issue 1, 1-9

Abstract: Abstract To ensure accurate DNA replication, a replisome must effectively overcome numerous obstacles on its DNA substrate. After encountering an obstacle, a progressing replisome often aborts DNA synthesis but continues to unwind. However, little is known about how DNA synthesis is resumed downstream of an obstacle. Here, we examine the consequences of a non-replicating replisome collision with a co-directional RNA polymerase (RNAP). Using single-molecule and ensemble methods, we find that T7 helicase interacts strongly with a non-replicating T7 DNA polymerase (DNAP) at a replication fork. As the helicase advances, the associated DNAP also moves forward. The presence of the DNAP increases both helicase’s processivity and unwinding rate. We show that such a DNAP, together with its helicase, is indeed able to actively disrupt a stalled transcription elongation complex, and then initiates replication using the RNA transcript as a primer. These observations exhibit T7 helicase’s novel role in replication re-initiation.

Date: 2018
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DOI: 10.1038/s41467-018-04702-x

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