PcrA-mediated disruption of RecA nucleoprotein filaments—essential role of the ATPase activity of RecA
Matthew V. Fagerburga, Grant D. Schauerb c, Karen R. Thickmanb c, Piero R. Biancob c, Saleem Ahmad Khanb c, Sanford H. Leubab c, Syam Prasad Anandb c✉
- aDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA
- bDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, 2 Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15219, 3 Department of Microbiology and Immunology, Center for Single Molecule Biophysics, University at Buffalo, Buffalo, NY 14214 and 4 University of Pittsburgh Cancer Institute, Pittsburgh, PA, 15213, USA
- cUniversity of Pittsburgh, Pittsburgh, United States
- ✉Corresponding author
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Abstract
The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a 'head-on collision' between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.
TL;DRIt is demonstrated that translocation of PcrA is not sufficient to displace RecA from the DNA and an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments is assigned. (Semantic Scholar)