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Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination

Human RAD52 promotes annealing of complementary single-stranded DNA (ssDNA). In-depth knowledge of RAD52-DNA interaction is required to understand how its activity is integrated in DNA repair processes. Here, we visualize individual fluorescent RAD52 complexes interacting with single DNA molecules....

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Autores principales: Brouwer, Ineke, Zhang, Hongshan, Candelli, Andrea, Normanno, Davide, Peterman, Erwin J.G., Wuite, Gijs J.L., Modesti, Mauro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5379009/
https://www.ncbi.nlm.nih.gov/pubmed/28329678
http://dx.doi.org/10.1016/j.celrep.2017.02.068
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author Brouwer, Ineke
Zhang, Hongshan
Candelli, Andrea
Normanno, Davide
Peterman, Erwin J.G.
Wuite, Gijs J.L.
Modesti, Mauro
author_facet Brouwer, Ineke
Zhang, Hongshan
Candelli, Andrea
Normanno, Davide
Peterman, Erwin J.G.
Wuite, Gijs J.L.
Modesti, Mauro
author_sort Brouwer, Ineke
collection PubMed
description Human RAD52 promotes annealing of complementary single-stranded DNA (ssDNA). In-depth knowledge of RAD52-DNA interaction is required to understand how its activity is integrated in DNA repair processes. Here, we visualize individual fluorescent RAD52 complexes interacting with single DNA molecules. The interaction with ssDNA is rapid, static, and tight, where ssDNA appears to wrap around RAD52 complexes that promote intra-molecular bridging. With double-stranded DNA (dsDNA), interaction is slower, weaker, and often diffusive. Interestingly, force spectroscopy experiments show that RAD52 alters the mechanics dsDNA by enhancing DNA flexibility and increasing DNA contour length, suggesting intercalation. RAD52 binding changes the nature of the overstretching transition of dsDNA and prevents DNA melting, which is advantageous for strand clamping during or after annealing. DNA-bound RAD52 is efficient at capturing ssDNA in trans. Together, these effects may help key steps in DNA repair, such as second-end capture during homologous recombination or strand annealing during RAD51-independent recombination reactions.
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spelling pubmed-53790092017-04-07 Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination Brouwer, Ineke Zhang, Hongshan Candelli, Andrea Normanno, Davide Peterman, Erwin J.G. Wuite, Gijs J.L. Modesti, Mauro Cell Rep Report Human RAD52 promotes annealing of complementary single-stranded DNA (ssDNA). In-depth knowledge of RAD52-DNA interaction is required to understand how its activity is integrated in DNA repair processes. Here, we visualize individual fluorescent RAD52 complexes interacting with single DNA molecules. The interaction with ssDNA is rapid, static, and tight, where ssDNA appears to wrap around RAD52 complexes that promote intra-molecular bridging. With double-stranded DNA (dsDNA), interaction is slower, weaker, and often diffusive. Interestingly, force spectroscopy experiments show that RAD52 alters the mechanics dsDNA by enhancing DNA flexibility and increasing DNA contour length, suggesting intercalation. RAD52 binding changes the nature of the overstretching transition of dsDNA and prevents DNA melting, which is advantageous for strand clamping during or after annealing. DNA-bound RAD52 is efficient at capturing ssDNA in trans. Together, these effects may help key steps in DNA repair, such as second-end capture during homologous recombination or strand annealing during RAD51-independent recombination reactions. Cell Press 2017-03-21 /pmc/articles/PMC5379009/ /pubmed/28329678 http://dx.doi.org/10.1016/j.celrep.2017.02.068 Text en © 2017 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Report
Brouwer, Ineke
Zhang, Hongshan
Candelli, Andrea
Normanno, Davide
Peterman, Erwin J.G.
Wuite, Gijs J.L.
Modesti, Mauro
Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title_full Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title_fullStr Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title_full_unstemmed Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title_short Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination
title_sort human rad52 captures and holds dna strands, increases dna flexibility, and prevents melting of duplex dna: implications for dna recombination
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5379009/
https://www.ncbi.nlm.nih.gov/pubmed/28329678
http://dx.doi.org/10.1016/j.celrep.2017.02.068
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