2021
Nature

RecA finds homologous DNA by reduced dimensionality search

Authors:

Wiktor, J., Gynnå, A. H., Leroy, P., Larsson, J., Coceano, G., Testa, I., & Elf, J.

Keywords:

DNA recombination, Homologous recombination

Abstract:

Homologous recombination is essential for the accurate repair of double-stranded DNA breaks (DSBs). Initially, the RecBCD complex resects the ends of the DSB into 3′ single-stranded DNA on which a RecA filament assembles. Next, the filament locates the homologous repair template on the sister chromosome. 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 homologues, we believe this model to be widely conserved across living organisms.