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Visualization of DNA Repair Proteins Interaction by Immunofluorescence
Published on: June 26, 2020
The cohesin-like RecN protein stimulates RecA-mediated recombinational repair of DNA double-strand breaks
Lee A Uranga1, Emigdio D Reyes1, Praveen L Patidar1
1Department of Chemistry and Biochemistry, New Mexico State University, P.O. Box 30001, MSC 3C, Las Cruces, New Mexico 88003, USA.
Abstract:
RecN is a cohesin-like protein involved in DNA double-strand break repair in bacteria. The RecA recombinase functions to mediate repair via homologous DNA strand invasion to form D-loops. Here we provide evidence that the RecN protein stimulates the DNA strand invasion step of RecA-mediated recombinational DNA repair. The intermolecular DNA tethering activity of RecN protein described previously cannot fully explain this novel activity since stimulation of RecA function is species-specific and requires RecN ATP hydrolysis. Further, DNA-bound RecA protein increases the rate of ATP hydrolysis catalysed by RecN during the DNA pairing reaction. DNA-dependent RecN ATPase kinetics are affected by RecA protein in a manner suggesting a specific order of protein-DNA assembly, with RecN acting after RecA binds DNA. We present a model for RecN function that includes presynaptic stimulation of the bacterial repair pathway perhaps by contributing to the RecA homology search before ternary complex formation.
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