High-resolution structure of the presynaptic RAD51 filament on single-stranded DNA by electron cryo-microscopy

Short, Judith M.; Liu, Yang; Chen, Shao-Xia; Soni, Neelesh; Madhusudhan, M.S.; Shivji, Mahmud K. K.; Venkitaraman, Ashok R.

doi:10.1093/nar/gkw783

Cited by 75 publications

(96 citation statements)

References 57 publications

(123 reference statements)

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“…Our reconstruction of the RAD51 presynaptic complex share a very similar overall shape of the protomer with the 3D reconstruction of HsRAD51 presynaptic complex reported in a latest research although the helical symmetry deviates slightly between the two. 38 …”

Section: Resultsmentioning

confidence: 99%

Cryo-EM structures of human RAD51 recombinase filaments during catalysis of DNA-strand exchange

Zhao

et al. 2016

Nat Struct Mol Biol

116

166

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The central step in eukaryotic homologous recombination (HR) is ATP-dependent DNA strand exchange mediated by the Rad51 recombinase. In this process, Rad51 assembles on single-stranded DNA (ssDNA) to yield a helical filament that is able to search for and invade a homologous double-stranded DNA (dsDNA), leading to strand separation in the dsDNA and formation of new base pairs between the initiating ssDNA and the complementary strand in the duplex partner. Here we have used cryo-electron microscopy (cryo-EM) to solve the structures of human RAD51 in complex with DNA molecules in their presynaptic and post-synaptic states at near atomic resolution. Our structures revealed both conserved and distinct structural features of the RAD51-DNA complexes in comparison with its prokaryotic counterpart. Importantly, we have also captured the structure of an arrested synaptic complex. The results provide insights into the molecular mechanism of DNA homology search and strand exchange processes.

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Section: Resultsmentioning

confidence: 99%

Cryo-EM structures of human RAD51 recombinase filaments during catalysis of DNA-strand exchange

Zhao

et al. 2016

Nat Struct Mol Biol

116

166

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“…More recently, high‐resolution cryo‐electron microscopy (cryoEM) studies of the presynaptic NPF in the presence of the ATP analogue AMP‐PNP have measured an average filament pitch of 10.3 nm with 6.4 protomers per turn (Short et al , 2016) and confirmed its structural polymorphism, with 80% of the filaments having pitch values in the range of 9.5–11.0 nm; a similar cryoEM analysis of the active form of the NPF reported a filament pitch of 10.0 nm with 6.3 protomers per turn (Xu et al , 2017). …”

Section: Introductionmentioning

confidence: 99%

Two distinct conformational states define the interaction of human RAD 51‐ ATP with single‐stranded DNA

et al. 2018

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An essential mechanism for repairing DNA double‐strand breaks is homologous recombination (HR). One of its core catalysts is human RAD51 (hRAD51), which assembles as a helical nucleoprotein filament on single‐stranded DNA, promoting DNA‐strand exchange. Here, we study the interaction of hRAD51 with single‐stranded DNA using a single‐molecule approach. We show that ATP‐bound hRAD51 filaments can exist in two different states with different contour lengths and with a free‐energy difference of ~4 kBT per hRAD51 monomer. Upon ATP hydrolysis, the filaments convert into a disassembly‐competent ADP‐bound configuration. In agreement with the single‐molecule analysis, we demonstrate the presence of two distinct protomer interfaces in the crystal structure of a hRAD51‐ATP filament, providing a structural basis for the two conformational states of the filament. Together, our findings provide evidence that hRAD51‐ATP filaments can exist in two interconvertible conformational states, which might be functionally relevant for DNA homology recognition and strand exchange.

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“…Although there were other differences in the proteins from these two studies, it is possible that the N-terminus constrains the filaments to a single conformation at the ATPase interface 8, 10 . More recently, high-resolution structural models of human RAD51 filaments have been obtained from cryo–electron microscopy reconstructions 11, 12 . Here, filaments formed with human RAD51 had only one conformation at the nucleotide-binding site.…”

Section: Rad51 Nucleoprotein Filaments: Variety In Form and Functionmentioning

confidence: 99%

Variation in RAD51 details a hub of functions: opportunities to advance cancer diagnosis and therapy

Zon¹,

Kanaar

Wyman³

2018

F1000Res

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Loss of genome stability is one of the hallmarks of the enabling characteristics of cancer development. Homologous recombination is a DNA repair process that often breaks down as a prelude to developing cancer. Conversely, homologous recombination can be the Achilles’ heel in common anti-cancer therapies, which are effective by inducing irreparable DNA damage. Here, we review recent structural and functional studies of RAD51, the protein that catalyzes the defining step of homologous recombination: homology recognition and DNA strand exchange. Specific mutations can be linked to structural changes and known essential functions. Additional RAD51 interactions and functions may be revealed. The identification of viable mutations in this essential protein may help define the range of activity and interactions needed. All of this information provides opportunities to fine-tune existing therapies based on homologous recombination status, guide diagnosis, and hopefully develop new clinical tools.

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High-resolution structure of the presynaptic RAD51 filament on single-stranded DNA by electron cryo-microscopy

Cited by 75 publications

References 57 publications

Cryo-EM structures of human RAD51 recombinase filaments during catalysis of DNA-strand exchange

Cryo-EM structures of human RAD51 recombinase filaments during catalysis of DNA-strand exchange

Two distinct conformational states define the interaction of human RAD 51‐ ATP with single‐stranded DNA

Variation in RAD51 details a hub of functions: opportunities to advance cancer diagnosis and therapy

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