RecA finds homologous DNA by reduced dimensionality search

Jakub Wiktor, Arvid H. Gynnå, Prune Leroy, Jimmy Larsson, Giovanna Coceano, Ilaria Testa and Johan Elf ()
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Jakub Wiktor: Uppsala University
Arvid H. Gynnå: Uppsala University
Prune Leroy: Uppsala University
Jimmy Larsson: Uppsala University
Giovanna Coceano: KTH Royal Institute of Technology
Ilaria Testa: KTH Royal Institute of Technology
Johan Elf: Uppsala University

Nature, 2021, vol. 597, issue 7876, 426-429

Abstract: Abstract Homologous recombination is essential for the accurate repair of double-stranded DNA breaks (DSBs)1. Initially, the RecBCD complex2 resects the ends of the DSB into 3′ single-stranded DNA on which a RecA filament assembles3. Next, the filament locates the homologous repair template on the sister chromosome4. Here we directly visualize the repair of DSBs in single cells, using high-throughput microfluidics and fluorescence microscopy. We find that, in Escherichia coli, repair of DSBs between segregated sister loci is completed in 15 ± 5 min (mean ± s.d.) with minimal fitness loss. We further show that the search takes less than 9 ± 3 min (mean ± s.d) and is mediated by a thin, highly dynamic RecA filament that stretches throughout the cell. We propose that the architecture of the RecA filament effectively reduces search dimensionality. This model predicts a search time that is consistent with our measurement and is corroborated by the observation that the search time does not depend on the length of the cell or the amount of DNA. Given the abundance of RecA homologues5, we believe this model to be widely conserved across living organisms.

Date: 2021
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DOI: 10.1038/s41586-021-03877-6

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