This study was supported by the National Science Foundation award MCB1916625 and by the National Institute of General Medical Sciences at the National Institutes of Health award R16GM145543.
Journal of Biological Chemistry
DNA replication -- Genomes
Accurately completing DNA replication when two forks converge is essential to genomic stability. The RecBCD helicase–nuclease complex plays a central role in completion by promoting resection and joining of the excess DNA created when replisomes converge. chi sequences alter RecBCD activity and localize with crossover hotspots during sexual events in bacteria, yet their functional role during chromosome replication remains unknown. Here, we use two-dimensional agarose gel analysis to show that chi induces replication on substrates containing convergent forks. The induced replication is processive but uncoupled with respect to leading and lagging strand synthesis and can be suppressed by ter sites which limit replisome progression. Our observations demonstrate that convergent replisomes create a substrate that is processed by RecBCD and that chi, when encountered, switches RecBCD from a degradative to replicative function. We propose that chi serves to functionally differentiate DNA ends created during completion, which require degradation, from those created by chromosomal double-strand breaks, which require resynthesis.
© 2023 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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Hamilton, N. A., Jehru, A. E., Samples, W. N., Wendel, B. M., Mokhtari, P. D., Courcelle, C. T., & Courcelle, J. (2023). chi sequences switch the RecBCD helicase–nuclease complex from degradative to replicative modes during the completion of DNA replication. Journal of Biological Chemistry, 299(3).