Human RAD51 rapidly forms intrinsically dynamic nucleoprotein filaments modulated by nucleotide binding state

Mário Spírek, Jarmila Mlčoušková, Ondrej Beláň, Máté Gyimesi, Gábor M. Harami, Eszter Molnár, Jiri Novacek, Mihály Kovács, Lumir Krejci

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Abstract

Formation of RAD51 filaments on single-stranded DNA is an essential event during homologous recombination, which is required for homology search, strand exchange and protection of replication forks. Formation of nucleoprotein filaments (NF) is required for development and genomic stability, and its failure is associated with developmental abnormalities and tumorigenesis. Here we describe the structure of the human RAD51 NFs and of its Walker box mutants using electron microscopy. Wild-type RAD51 filaments adopt an 'open' conformation when compared to a 'closed' structure formed by mutants, reflecting alterations in helical pitch. The kinetics of formation/disassembly of RAD51 filaments show rapid and high ssDNA coverage via low coopera-tivity binding of RAD51 units along the DNA. Subsequently, a series of isomerization or dissociation events mediated by nucleotide binding state creates intrinsically dynamic RAD51 NFs. Our findings highlight important a mechanistic divergence among recombinases from different organisms, in line with the diversity of biological mechanisms of HR initiation and quality control. These data reveal unexpected intrinsic dynamic properties of the RAD51 filament during assembly/disassembly, which may be important for the proper control of homologous recombination.

Original languageEnglish
Pages (from-to)3967-3980
Number of pages14
JournalNucleic acids research
Volume46
Issue number8
DOIs
Publication statusPublished - Jan 1 2018

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ASJC Scopus subject areas

  • Genetics

Cite this

Spírek, M., Mlčoušková, J., Beláň, O., Gyimesi, M., Harami, G. M., Molnár, E., Novacek, J., Kovács, M., & Krejci, L. (2018). Human RAD51 rapidly forms intrinsically dynamic nucleoprotein filaments modulated by nucleotide binding state. Nucleic acids research, 46(8), 3967-3980. https://doi.org/10.1093/nar/gky111