T7 replisome directly overcomes DNA damage

Bo Sun, Manjula Pandey, James T. Inman, Yi Yang, Mikhail Kashlev, Smita S. Patel and Michelle D. Wang ()
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Bo Sun: Laboratory of Atomic and Solid State Physics, Cornell University
Manjula Pandey: Rutgers-Robert Wood Johnson Medical School
James T. Inman: Laboratory of Atomic and Solid State Physics, Cornell University
Yi Yang: Laboratory of Atomic and Solid State Physics, Cornell University
Mikhail Kashlev: NCI Center for Cancer Research
Smita S. Patel: Rutgers-Robert Wood Johnson Medical School
Michelle D. Wang: Laboratory of Atomic and Solid State Physics, Cornell University

Nature Communications, 2015, vol. 6, issue 1, 1-8

Abstract: Abstract Cells and viruses possess several known ‘restart’ pathways to overcome lesions during DNA replication. However, these ‘bypass’ pathways leave a gap in replicated DNA or require recruitment of accessory proteins, resulting in significant delays to fork movement or even cell division arrest. Using single-molecule and ensemble methods, we demonstrate that the bacteriophage T7 replisome is able to directly replicate through a leading-strand cyclobutane pyrimidine dimer (CPD) lesion. We show that when a replisome encounters the lesion, a substantial fraction of DNA polymerase (DNAP) and helicase stay together at the lesion, the replisome does not dissociate and the helicase does not move forward on its own. The DNAP is able to directly replicate through the lesion by working in conjunction with helicase through specific helicase–DNAP interactions. These observations suggest that the T7 replisome is fundamentally permissive of DNA lesions via pathways that do not require fork adjustment or replisome reassembly.

Date: 2015
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DOI: 10.1038/ncomms10260

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