Srs2 prevents Rad51 filament formation by repetitive motion on DNA

Yupeng Qiu; Edwin Antony; Sultan Doganay; Hye Ran Koh; Timothy M. Lohman; Sua Myong

doi:10.1038/ncomms3281

Srs2 prevents Rad51 filament formation by repetitive motion on DNA

Yupeng Qiu, Edwin Antony, Sultan Doganay, Hye Ran Koh, Timothy M. Lohman, Sua Myong

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Abstract

Srs2 dismantles presynaptic Rad51 filaments and prevents its re-formation as an anti-recombinase. However, the molecular mechanism by which Srs2 accomplishes these tasks remains unclear. Here we report a single-molecule fluorescence study of the dynamics of Rad51 filament formation and its disruption by Srs2. Rad51 forms filaments on single-stranded DNA by sequential binding of primarily monomers and dimers in a 5′-3′ direction. One Rad51 molecule binds to three nucleotides, and six monomers are required to achieve a stable nucleation cluster. Srs2 exhibits ATP-dependent repetitive motion on single-stranded DNA and this activity prevents re-formation of the Rad51 filament. The same activity of Srs2 cannot prevent RecA filament formation, indicating its specificity for Rad51. Srs2's DNA-unwinding activity is greatly suppressed when Rad51 filaments form on duplex DNA. Taken together, our results reveal an exquisite and highly specific mechanism by which Srs2 regulates the Rad51 filament formation.

Original language	English
Article number	2281
Journal	Nature communications
Volume	4
DOIs	https://doi.org/10.1038/ncomms3281
State	Published - 2013

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@article{273deedf1c6a49668ad2ca9d5c5c1a03,

title = "Srs2 prevents Rad51 filament formation by repetitive motion on DNA",

abstract = "Srs2 dismantles presynaptic Rad51 filaments and prevents its re-formation as an anti-recombinase. However, the molecular mechanism by which Srs2 accomplishes these tasks remains unclear. Here we report a single-molecule fluorescence study of the dynamics of Rad51 filament formation and its disruption by Srs2. Rad51 forms filaments on single-stranded DNA by sequential binding of primarily monomers and dimers in a 5′-3′ direction. One Rad51 molecule binds to three nucleotides, and six monomers are required to achieve a stable nucleation cluster. Srs2 exhibits ATP-dependent repetitive motion on single-stranded DNA and this activity prevents re-formation of the Rad51 filament. The same activity of Srs2 cannot prevent RecA filament formation, indicating its specificity for Rad51. Srs2's DNA-unwinding activity is greatly suppressed when Rad51 filaments form on duplex DNA. Taken together, our results reveal an exquisite and highly specific mechanism by which Srs2 regulates the Rad51 filament formation.",

author = "Yupeng Qiu and Edwin Antony and Sultan Doganay and {Ran Koh}, Hye and Lohman, {Timothy M.} and Sua Myong",

note = "Funding Information: We thank members of the Myong laboratory for helpful discussions. We also thank Paul Hergenrother and Robert Tamp{\'e} for providing biotin–tris–NTA used for Srs2 tethering and NTA–Atto 647 used for Srs2 labelling, respectively. This work was supported by American Cancer Society (Research Scholar Grant; RSG-12-066-01-DMC), NIH Director{\textquoteright}s New Innovator Award (1DP2GM105453), the Human Frontier Science Program (RGP0007/2012) and the U.S. National Science Foundation Physics Frontiers Center Program (0822613) through the Center for the Physics of Living Cells for S.M. and NIH GM045948 for T.M.L.",

year = "2013",

doi = "10.1038/ncomms3281",

language = "English",

volume = "4",

journal = "Nature communications",

issn = "2041-1723",

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T1 - Srs2 prevents Rad51 filament formation by repetitive motion on DNA

AU - Qiu, Yupeng

AU - Antony, Edwin

AU - Doganay, Sultan

AU - Ran Koh, Hye

AU - Lohman, Timothy M.

AU - Myong, Sua

N1 - Funding Information: We thank members of the Myong laboratory for helpful discussions. We also thank Paul Hergenrother and Robert Tampé for providing biotin–tris–NTA used for Srs2 tethering and NTA–Atto 647 used for Srs2 labelling, respectively. This work was supported by American Cancer Society (Research Scholar Grant; RSG-12-066-01-DMC), NIH Director’s New Innovator Award (1DP2GM105453), the Human Frontier Science Program (RGP0007/2012) and the U.S. National Science Foundation Physics Frontiers Center Program (0822613) through the Center for the Physics of Living Cells for S.M. and NIH GM045948 for T.M.L.

PY - 2013

Y1 - 2013

N2 - Srs2 dismantles presynaptic Rad51 filaments and prevents its re-formation as an anti-recombinase. However, the molecular mechanism by which Srs2 accomplishes these tasks remains unclear. Here we report a single-molecule fluorescence study of the dynamics of Rad51 filament formation and its disruption by Srs2. Rad51 forms filaments on single-stranded DNA by sequential binding of primarily monomers and dimers in a 5′-3′ direction. One Rad51 molecule binds to three nucleotides, and six monomers are required to achieve a stable nucleation cluster. Srs2 exhibits ATP-dependent repetitive motion on single-stranded DNA and this activity prevents re-formation of the Rad51 filament. The same activity of Srs2 cannot prevent RecA filament formation, indicating its specificity for Rad51. Srs2's DNA-unwinding activity is greatly suppressed when Rad51 filaments form on duplex DNA. Taken together, our results reveal an exquisite and highly specific mechanism by which Srs2 regulates the Rad51 filament formation.

AB - Srs2 dismantles presynaptic Rad51 filaments and prevents its re-formation as an anti-recombinase. However, the molecular mechanism by which Srs2 accomplishes these tasks remains unclear. Here we report a single-molecule fluorescence study of the dynamics of Rad51 filament formation and its disruption by Srs2. Rad51 forms filaments on single-stranded DNA by sequential binding of primarily monomers and dimers in a 5′-3′ direction. One Rad51 molecule binds to three nucleotides, and six monomers are required to achieve a stable nucleation cluster. Srs2 exhibits ATP-dependent repetitive motion on single-stranded DNA and this activity prevents re-formation of the Rad51 filament. The same activity of Srs2 cannot prevent RecA filament formation, indicating its specificity for Rad51. Srs2's DNA-unwinding activity is greatly suppressed when Rad51 filaments form on duplex DNA. Taken together, our results reveal an exquisite and highly specific mechanism by which Srs2 regulates the Rad51 filament formation.

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U2 - 10.1038/ncomms3281

DO - 10.1038/ncomms3281

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SN - 2041-1723

VL - 4

JO - Nature communications

JF - Nature communications

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Srs2 prevents Rad51 filament formation by repetitive motion on DNA

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