TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity

Pavel Moudry; Kenji Watanabe; Kamila M. Wolanin; Jirina Bartkova; Isabel E. Wassing; Sugiko Watanabe; Robert Strauss; Rune Troelsgaard Pedersen; Vibe Hallundbæk Østergaard; Michael Lisby; Miguel Andújar-Sánchez; Apolinar Maya-Mendoza; Fumiko Esashi; Jiri Lukas; Jiri Bartek

doi:10.1083/jcb.201507042

TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity

Pavel Moudry, Kenji Watanabe, Kamila M. Wolanin, Jirina Bartkova, Isabel E. Wassing, Sugiko Watanabe, Robert Strauss, Rune Troelsgaard Pedersen, Vibe Hallundbæk Østergaard, Michael Lisby, Miguel Andújar-Sánchez, Apolinar Maya-Mendoza, Fumiko Esashi, Jiri Lukas, Jiri Bartek

Functional Genomics

34 Citations (Scopus)

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Abstract

Topoisomerase IIβ-binding protein 1 (TOPBP1) participates in DNA replication and DNA damage response; however, its role in DNA repair and relevance for human cancer remain unclear. Here, through an unbiased small interfering RNA screen, we identified and validated TOPBP1 as a novel determinant whose loss sensitized human cells to olaparib, an inhibitor of poly(ADP-ribose) polymerase. We show that TOPBP1 acts in homologous recombination (HR) repair, impacts olaparib response, and exhibits aberrant patterns in subsets of human ovarian carcinomas. TOPBP1 depletion abrogated RAD51 loading to chromatin and formation of RAD51 foci, but without affecting the upstream HR steps of DNA end resection and RPA loading. Furthermore, TOPBP1 BRCT domains 7/8 are essential for RAD51 foci formation. Mechanistically, TOPBP1 physically binds PLK1 and promotes PLK1 kinase-mediated phosphorylation of RAD51 at serine 14, a modification required for RAD51 recruitment to chromatin. Overall, our results provide mechanistic insights into TOPBP1's role in HR, with potential clinical implications for cancer treatment.

Original language	English
Journal	The Journal of Cell Biology
Volume	212
Issue number	3
Pages (from-to)	281-288
Number of pages	8
ISSN	0021-9525
DOIs	https://doi.org/10.1083/jcb.201507042
Publication status	Published - 2016

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Moudry, P., Watanabe, K., Wolanin, K. M., Bartkova, J., Wassing, I. E., Watanabe, S., Strauss, R., Pedersen, R. T., Østergaard, V. H., Lisby, M., Andújar-Sánchez, M., Maya-Mendoza, A., Esashi, F., Lukas, J., & Bartek, J. (2016). TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity. The Journal of Cell Biology, 212(3), 281-288. https://doi.org/10.1083/jcb.201507042

Moudry, P, Watanabe, K, Wolanin, KM, Bartkova, J, Wassing, IE, Watanabe, S, Strauss, R, Pedersen, RT, Østergaard, VH , Lisby, M, Andújar-Sánchez, M, Maya-Mendoza, A, Esashi, F, Lukas, J & Bartek, J 2016, 'TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity', The Journal of Cell Biology, vol. 212, no. 3, pp. 281-288. https://doi.org/10.1083/jcb.201507042

@article{ff52603e126f47ef88b73d94520e3d94,

title = "TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity",

abstract = "Topoisomerase IIβ-binding protein 1 (TOPBP1) participates in DNA replication and DNA damage response; however, its role in DNA repair and relevance for human cancer remain unclear. Here, through an unbiased small interfering RNA screen, we identified and validated TOPBP1 as a novel determinant whose loss sensitized human cells to olaparib, an inhibitor of poly(ADP-ribose) polymerase. We show that TOPBP1 acts in homologous recombination (HR) repair, impacts olaparib response, and exhibits aberrant patterns in subsets of human ovarian carcinomas. TOPBP1 depletion abrogated RAD51 loading to chromatin and formation of RAD51 foci, but without affecting the upstream HR steps of DNA end resection and RPA loading. Furthermore, TOPBP1 BRCT domains 7/8 are essential for RAD51 foci formation. Mechanistically, TOPBP1 physically binds PLK1 and promotes PLK1 kinase-mediated phosphorylation of RAD51 at serine 14, a modification required for RAD51 recruitment to chromatin. Overall, our results provide mechanistic insights into TOPBP1's role in HR, with potential clinical implications for cancer treatment.",

author = "Pavel Moudry and Kenji Watanabe and Wolanin, {Kamila M.} and Jirina Bartkova and Wassing, {Isabel E.} and Sugiko Watanabe and Robert Strauss and Pedersen, {Rune Troelsgaard} and {\O}stergaard, {Vibe Hallundb{\ae}k} and Michael Lisby and Miguel And{\'u}jar-S{\'a}nchez and Apolinar Maya-Mendoza and Fumiko Esashi and Jiri Lukas and Jiri Bartek",

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doi = "10.1083/jcb.201507042",

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T1 - TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity

AU - Moudry, Pavel

AU - Watanabe, Kenji

AU - Wolanin, Kamila M.

AU - Bartkova, Jirina

AU - Wassing, Isabel E.

AU - Watanabe, Sugiko

AU - Strauss, Robert

AU - Pedersen, Rune Troelsgaard

AU - Østergaard, Vibe Hallundbæk

AU - Lisby, Michael

AU - Andújar-Sánchez, Miguel

AU - Maya-Mendoza, Apolinar

AU - Esashi, Fumiko

AU - Lukas, Jiri

AU - Bartek, Jiri

PY - 2016

Y1 - 2016

N2 - Topoisomerase IIβ-binding protein 1 (TOPBP1) participates in DNA replication and DNA damage response; however, its role in DNA repair and relevance for human cancer remain unclear. Here, through an unbiased small interfering RNA screen, we identified and validated TOPBP1 as a novel determinant whose loss sensitized human cells to olaparib, an inhibitor of poly(ADP-ribose) polymerase. We show that TOPBP1 acts in homologous recombination (HR) repair, impacts olaparib response, and exhibits aberrant patterns in subsets of human ovarian carcinomas. TOPBP1 depletion abrogated RAD51 loading to chromatin and formation of RAD51 foci, but without affecting the upstream HR steps of DNA end resection and RPA loading. Furthermore, TOPBP1 BRCT domains 7/8 are essential for RAD51 foci formation. Mechanistically, TOPBP1 physically binds PLK1 and promotes PLK1 kinase-mediated phosphorylation of RAD51 at serine 14, a modification required for RAD51 recruitment to chromatin. Overall, our results provide mechanistic insights into TOPBP1's role in HR, with potential clinical implications for cancer treatment.

AB - Topoisomerase IIβ-binding protein 1 (TOPBP1) participates in DNA replication and DNA damage response; however, its role in DNA repair and relevance for human cancer remain unclear. Here, through an unbiased small interfering RNA screen, we identified and validated TOPBP1 as a novel determinant whose loss sensitized human cells to olaparib, an inhibitor of poly(ADP-ribose) polymerase. We show that TOPBP1 acts in homologous recombination (HR) repair, impacts olaparib response, and exhibits aberrant patterns in subsets of human ovarian carcinomas. TOPBP1 depletion abrogated RAD51 loading to chromatin and formation of RAD51 foci, but without affecting the upstream HR steps of DNA end resection and RPA loading. Furthermore, TOPBP1 BRCT domains 7/8 are essential for RAD51 foci formation. Mechanistically, TOPBP1 physically binds PLK1 and promotes PLK1 kinase-mediated phosphorylation of RAD51 at serine 14, a modification required for RAD51 recruitment to chromatin. Overall, our results provide mechanistic insights into TOPBP1's role in HR, with potential clinical implications for cancer treatment.

U2 - 10.1083/jcb.201507042

DO - 10.1083/jcb.201507042

M3 - Journal article

C2 - 26811421

SN - 0021-9525

VL - 212

SP - 281

EP - 288

JO - Journal of Cell Biology

JF - Journal of Cell Biology

IS - 3

ER -

TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity

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