A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination

Anderson T. Wang; Taeho Kim; John E. Wagner; Brooke A. Conti; Francis P. Lach; Athena L. Huang; Henrik Molina; Erica M. Sanborn; Heather Zierhut; Belinda K. Cornes; Avinash Abhyankar; Carrie Sougnez; Stacey B. Gabriel; Arleen D. Auerbach; Stephen C. Kowalczykowski; Agata Smogorzewska

doi:10.1016/j.molcel.2015.07.009

A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination

Anderson T. Wang, Taeho Kim, John E. Wagner, Brooke A. Conti, Francis P. Lach, Athena L. Huang, Henrik Molina, Erica M. Sanborn, Heather Zierhut, Belinda K. Cornes, Avinash Abhyankar, Carrie Sougnez, Stacey B. Gabriel, Arleen D. Auerbach, Stephen C. Kowalczykowski, Agata Smogorzewska

Research output: Contribution to journal › Article › peer-review

144 Scopus citations

Abstract

Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. Invitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio ofmutantto wild-type RAD51 in cells. Instead, patientcells aresensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity.

Original language	English (US)
Pages (from-to)	478-490
Number of pages	13
Journal	Molecular Cell
Volume	59
Issue number	3
DOIs	https://doi.org/10.1016/j.molcel.2015.07.009
State	Published - Aug 6 2015

Bibliographical note

Funding Information:
We thank the patient and her family, without whom this study would not be possible. We thank Steve West and John Petrini for providing us with anti-RAD51 and anti-MRE11 antibodies, respectively; Nikola Pavletich for the RAD51 cDNA construct; and Detlev Schindler for sharing the RAD50 mutant cell line. We are grateful to all members of the A.S. laboratory for helpful discussion and critical comments on the manuscript. This work was supported by a Starr Center Consortium grant (A.S. and S.B.G.), the Rita Allen Foundation and Burroughs Wellcome Foundation (A.S.), an Irma T. Hirschl Research Award (A.S.), a Doris Duke Clinical Scientist Development Award (A.S.), NIH grant GM62653 (S.C.K.), grant #8 UL1 TR000043 from the National Center for Advancing Translational Sciences (NCATS), and the NIH Clinical and Translational Science Award (CTSA) program. The IFAR is partially supported by NIH grant R01HL120922 (A.S.). A.T.W. was partially funded by Congressionally Directed Medical Research Programs and a Bone Marrow Failure Research Program Postdoctoral Fellowship Training Award (grant log #BM120004).

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© 2015 Elsevier Inc.

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Wang, A. T., Kim, T., Wagner, J. E., Conti, B. A., Lach, F. P., Huang, A. L., Molina, H., Sanborn, E. M., Zierhut, H., Cornes, B. K., Abhyankar, A., Sougnez, C., Gabriel, S. B., Auerbach, A. D., Kowalczykowski, S. C., & Smogorzewska, A. (2015). A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination. Molecular Cell, 59(3), 478-490. https://doi.org/10.1016/j.molcel.2015.07.009

A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination. / Wang, Anderson T.; Kim, Taeho; Wagner, John E.; Conti, Brooke A.; Lach, Francis P.; Huang, Athena L.; Molina, Henrik; Sanborn, Erica M.; Zierhut, Heather; Cornes, Belinda K.; Abhyankar, Avinash; Sougnez, Carrie; Gabriel, Stacey B.; Auerbach, Arleen D.; Kowalczykowski, Stephen C.; Smogorzewska, Agata.

In: Molecular Cell, Vol. 59, No. 3, 06.08.2015, p. 478-490.

Research output: Contribution to journal › Article › peer-review

Wang, AT, Kim, T, Wagner, JE, Conti, BA, Lach, FP, Huang, AL, Molina, H, Sanborn, EM, Zierhut, H, Cornes, BK, Abhyankar, A, Sougnez, C, Gabriel, SB, Auerbach, AD, Kowalczykowski, SC & Smogorzewska, A 2015, 'A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination', Molecular Cell, vol. 59, no. 3, pp. 478-490. https://doi.org/10.1016/j.molcel.2015.07.009

Wang, Anderson T. ; Kim, Taeho ; Wagner, John E. ; Conti, Brooke A. ; Lach, Francis P. ; Huang, Athena L. ; Molina, Henrik ; Sanborn, Erica M. ; Zierhut, Heather ; Cornes, Belinda K. ; Abhyankar, Avinash ; Sougnez, Carrie ; Gabriel, Stacey B. ; Auerbach, Arleen D. ; Kowalczykowski, Stephen C. ; Smogorzewska, Agata. / A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination. In: Molecular Cell. 2015 ; Vol. 59, No. 3. pp. 478-490.

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title = "A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination",

abstract = "Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. Invitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio ofmutantto wild-type RAD51 in cells. Instead, patientcells aresensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity.",

author = "Wang, {Anderson T.} and Taeho Kim and Wagner, {John E.} and Conti, {Brooke A.} and Lach, {Francis P.} and Huang, {Athena L.} and Henrik Molina and Sanborn, {Erica M.} and Heather Zierhut and Cornes, {Belinda K.} and Avinash Abhyankar and Carrie Sougnez and Gabriel, {Stacey B.} and Auerbach, {Arleen D.} and Kowalczykowski, {Stephen C.} and Agata Smogorzewska",

note = "Funding Information: We thank the patient and her family, without whom this study would not be possible. We thank Steve West and John Petrini for providing us with anti-RAD51 and anti-MRE11 antibodies, respectively; Nikola Pavletich for the RAD51 cDNA construct; and Detlev Schindler for sharing the RAD50 mutant cell line. We are grateful to all members of the A.S. laboratory for helpful discussion and critical comments on the manuscript. This work was supported by a Starr Center Consortium grant (A.S. and S.B.G.), the Rita Allen Foundation and Burroughs Wellcome Foundation (A.S.), an Irma T. Hirschl Research Award (A.S.), a Doris Duke Clinical Scientist Development Award (A.S.), NIH grant GM62653 (S.C.K.), grant #8 UL1 TR000043 from the National Center for Advancing Translational Sciences (NCATS), and the NIH Clinical and Translational Science Award (CTSA) program. The IFAR is partially supported by NIH grant R01HL120922 (A.S.). A.T.W. was partially funded by Congressionally Directed Medical Research Programs and a Bone Marrow Failure Research Program Postdoctoral Fellowship Training Award (grant log #BM120004). Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

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AU - Kim, Taeho

AU - Wagner, John E.

AU - Conti, Brooke A.

AU - Lach, Francis P.

AU - Huang, Athena L.

AU - Molina, Henrik

AU - Sanborn, Erica M.

AU - Zierhut, Heather

AU - Cornes, Belinda K.

AU - Abhyankar, Avinash

AU - Sougnez, Carrie

AU - Gabriel, Stacey B.

AU - Auerbach, Arleen D.

AU - Kowalczykowski, Stephen C.

AU - Smogorzewska, Agata

N1 - Funding Information: We thank the patient and her family, without whom this study would not be possible. We thank Steve West and John Petrini for providing us with anti-RAD51 and anti-MRE11 antibodies, respectively; Nikola Pavletich for the RAD51 cDNA construct; and Detlev Schindler for sharing the RAD50 mutant cell line. We are grateful to all members of the A.S. laboratory for helpful discussion and critical comments on the manuscript. This work was supported by a Starr Center Consortium grant (A.S. and S.B.G.), the Rita Allen Foundation and Burroughs Wellcome Foundation (A.S.), an Irma T. Hirschl Research Award (A.S.), a Doris Duke Clinical Scientist Development Award (A.S.), NIH grant GM62653 (S.C.K.), grant #8 UL1 TR000043 from the National Center for Advancing Translational Sciences (NCATS), and the NIH Clinical and Translational Science Award (CTSA) program. The IFAR is partially supported by NIH grant R01HL120922 (A.S.). A.T.W. was partially funded by Congressionally Directed Medical Research Programs and a Bone Marrow Failure Research Program Postdoctoral Fellowship Training Award (grant log #BM120004). Publisher Copyright: © 2015 Elsevier Inc.

PY - 2015/8/6

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N2 - Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. Invitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio ofmutantto wild-type RAD51 in cells. Instead, patientcells aresensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity.

AB - Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. Invitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio ofmutantto wild-type RAD51 in cells. Instead, patientcells aresensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity.

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A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination

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