Nucleotide proofreading functions by nematode RAD51 paralogs facilitate optimal RAD51 filament function

ŠPÍREK, Mário, M. R. G. TAYLOR, Ondrej BELÁŇ, S. J. BOULTON and Lumír KREJČÍ. Nucleotide proofreading functions by nematode RAD51 paralogs facilitate optimal RAD51 filament function. Nature Communications. London: Nature Publishing Group, 2021, vol. 12, No 1, p. 1-12. ISSN 2041-1723. Available from: https://dx.doi.org/10.1038/s41467-021-25830-x.

Basic information

• Original name: Nucleotide proofreading functions by nematode RAD51 paralogs facilitate optimal RAD51 filament function
• Authors: ŠPÍREK, Mário (703 Slovakia, belonging to the institution), M. R. G. TAYLOR, Ondrej BELÁŇ (703 Slovakia, belonging to the institution), S. J. BOULTON and Lumír KREJČÍ (203 Czech Republic, guarantor, belonging to the institution).
• Edition: Nature Communications, London, Nature Publishing Group, 2021, 2041-1723.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10608 Biochemistry and molecular biology
• Country of publisher: Germany
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 17.694
• RIV identification code: RIV/00216224:14110/21:00119274
• Organization unit: Faculty of Medicine
• Doi: http://dx.doi.org/10.1038/s41467-021-25830-x
• UT WoS: 000698606100024
• Keywords in English: RAD51; nucleotide proofreading functions
• Tags: 14110513, podil, rivok
• Tags: International impact, Reviewed

Abstract

The RAD51 recombinase assembles as helical nucleoprotein filaments on single-stranded DNA (ssDNA) and mediates invasion and strand exchange with homologous duplex DNA (dsDNA) during homologous recombination (HR), as well as protection and restart of stalled replication forks. Strand invasion by RAD51-ssDNA complexes depends on ATP binding. However, RAD51 can bind ssDNA in non-productive ADP-bound or nucleotide-free states, and ATP-RAD51-ssDNA complexes hydrolyse ATP over time. Here, we define unappreciated mechanisms by which the RAD51 paralog complex RFS-1/RIP-1 limits the accumulation of RAD-51-ssDNA complexes with unfavorable nucleotide content. We find RAD51 paralogs promote the turnover of ADP-bound RAD-51 from ssDNA, in striking contrast to their ability to stabilize productive ATP-bound RAD-51 nucleoprotein filaments. In addition, RFS-1/RIP-1 inhibits binding of nucleotide-free RAD-51 to ssDNA. We propose that 'nucleotide proofreading' activities of RAD51 paralogs co-operate to ensure the enrichment of active, ATP-bound RAD-51 filaments on ssDNA to promote HR. A RAD51 paralog complex, RFS-1/RIP-1, is shown to control ssDNA binding and dissociation by RAD-51 differentially in the presence and absence of nucleotide cofactors. These nucleotide proofreading activities drive a preferential accumulation of RAD-51-ssDNA complexes with optimal nucleotide content.

Links

• EF16_025/0007381, research and development project: Name: Preklinická progrese nových organických sloučenin s cílenou biologickou aktivitou
• GX21-22593X, research and development project: Name: Identifikace a charakterizace proteinů zahrnutých v metabolismu G-kvadruplexů a R-smyček a jejich vztah k replikaci DNA
• MUNI/G/1594/2019, interní kód MU: Name: RecQ4 – a protein hub required for proper replication and recombination and its implications in Rothmund-Thomson Syndrome

Investor: Masaryk University, INTERDISCIPLINARY - Interdisciplinary research projects

• 206292/E/17/Z, interní kód MU: Name: Mechanics and execution of homologous recombination - biophysics to the organism

Investor: Wellcome Trust