A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination

Guang Yu Zhao; Eiichiro Sonoda; Louise J. Barber; Hayato Oka; Yasuhiro Murakawa; Kouichi Yamada; Tsuyoshi Ikura; Xin Wang; Masahiko Kobayashi; Kenichi Yamamoto; Simon J. Boulton; Shunichi Takeda

doi:10.1016/j.molcel.2007.01.029

A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination

Guang Yu Zhao, Eiichiro Sonoda, Louise J. Barber, Hayato Oka, Yasuhiro Murakawa, Kouichi Yamada, Tsuyoshi Ikura, Xin Wang, Masahiko Kobayashi, Kenichi Yamamoto, Simon J. Boulton, Shunichi Takeda

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

190 Citations (Scopus)

Abstract

The ubiquitin (Ub)-conjugating enzyme Ubc13 is implicated in Rad6/Rad18-dependent postreplication repair (PRR) in budding yeast, but its function in vertebrates is not known. We show here that disruption or siRNA depletion of UBC13 in chicken DT40 or human cells confers severe growth defects due to chromosome instability, and hypersensitivity to both UV and ionizing radiation, consistent with a conserved role for Ubc13 in PRR. Remarkably, Ubc13-deficient cells are also compromised for DNA double-strand break (DSB) repair by homologous recombination (HR). Recruitment and activation of the E3 Ub ligase function of BRCA1 and the subsequent formation of the Rad51 nucleoprotein filament at DSBs are abolished in Ubc13-deficient cells. Furthermore, generation of ssDNA/RPA complexes at DSBs is severely attenuated in the absence of Ubc13. These data reveal a critical and unexpected role for vertebrate Ubc13 in the initiation of HR at the level of DSB processing.

Original language	English
Pages (from-to)	663-675
Number of pages	13
Journal	Molecular Cell
Volume	25
Issue number	5
DOIs	https://doi.org/10.1016/j.molcel.2007.01.029
Publication status	Published - Mar 9 2007
Externally published	Yes

Keywords

DNA
PROTEINS

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2007.01.029

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@article{cac446337e3e412385e5ac571571f05b,

title = "A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination",

abstract = "The ubiquitin (Ub)-conjugating enzyme Ubc13 is implicated in Rad6/Rad18-dependent postreplication repair (PRR) in budding yeast, but its function in vertebrates is not known. We show here that disruption or siRNA depletion of UBC13 in chicken DT40 or human cells confers severe growth defects due to chromosome instability, and hypersensitivity to both UV and ionizing radiation, consistent with a conserved role for Ubc13 in PRR. Remarkably, Ubc13-deficient cells are also compromised for DNA double-strand break (DSB) repair by homologous recombination (HR). Recruitment and activation of the E3 Ub ligase function of BRCA1 and the subsequent formation of the Rad51 nucleoprotein filament at DSBs are abolished in Ubc13-deficient cells. Furthermore, generation of ssDNA/RPA complexes at DSBs is severely attenuated in the absence of Ubc13. These data reveal a critical and unexpected role for vertebrate Ubc13 in the initiation of HR at the level of DSB processing.",

keywords = "DNA, PROTEINS",

author = "Zhao, {Guang Yu} and Eiichiro Sonoda and Barber, {Louise J.} and Hayato Oka and Yasuhiro Murakawa and Kouichi Yamada and Tsuyoshi Ikura and Xin Wang and Masahiko Kobayashi and Kenichi Yamamoto and Boulton, {Simon J.} and Shunichi Takeda",

note = "Funding Information: We thank Dr. Kimura (Nuclear Function and Dynamics Unit, HMRO, Kyoto University) for his help for microscopy analysis and histone purification. Financial support was provided in part by CREST.JST. (Saitama, Japan) and the center of excellence (COE) grant for Scientific Research from the Ministry of Education, Culture, Sports and Technology. L.J.B. and S.J.B. are funded by Cancer Research UK. ",

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doi = "10.1016/j.molcel.2007.01.029",

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journal = "Molecular Cell",

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T1 - A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination

AU - Zhao, Guang Yu

AU - Sonoda, Eiichiro

AU - Barber, Louise J.

AU - Oka, Hayato

AU - Murakawa, Yasuhiro

AU - Yamada, Kouichi

AU - Ikura, Tsuyoshi

AU - Wang, Xin

AU - Kobayashi, Masahiko

AU - Yamamoto, Kenichi

AU - Boulton, Simon J.

AU - Takeda, Shunichi

N1 - Funding Information: We thank Dr. Kimura (Nuclear Function and Dynamics Unit, HMRO, Kyoto University) for his help for microscopy analysis and histone purification. Financial support was provided in part by CREST.JST. (Saitama, Japan) and the center of excellence (COE) grant for Scientific Research from the Ministry of Education, Culture, Sports and Technology. L.J.B. and S.J.B. are funded by Cancer Research UK.

PY - 2007/3/9

Y1 - 2007/3/9

N2 - The ubiquitin (Ub)-conjugating enzyme Ubc13 is implicated in Rad6/Rad18-dependent postreplication repair (PRR) in budding yeast, but its function in vertebrates is not known. We show here that disruption or siRNA depletion of UBC13 in chicken DT40 or human cells confers severe growth defects due to chromosome instability, and hypersensitivity to both UV and ionizing radiation, consistent with a conserved role for Ubc13 in PRR. Remarkably, Ubc13-deficient cells are also compromised for DNA double-strand break (DSB) repair by homologous recombination (HR). Recruitment and activation of the E3 Ub ligase function of BRCA1 and the subsequent formation of the Rad51 nucleoprotein filament at DSBs are abolished in Ubc13-deficient cells. Furthermore, generation of ssDNA/RPA complexes at DSBs is severely attenuated in the absence of Ubc13. These data reveal a critical and unexpected role for vertebrate Ubc13 in the initiation of HR at the level of DSB processing.

AB - The ubiquitin (Ub)-conjugating enzyme Ubc13 is implicated in Rad6/Rad18-dependent postreplication repair (PRR) in budding yeast, but its function in vertebrates is not known. We show here that disruption or siRNA depletion of UBC13 in chicken DT40 or human cells confers severe growth defects due to chromosome instability, and hypersensitivity to both UV and ionizing radiation, consistent with a conserved role for Ubc13 in PRR. Remarkably, Ubc13-deficient cells are also compromised for DNA double-strand break (DSB) repair by homologous recombination (HR). Recruitment and activation of the E3 Ub ligase function of BRCA1 and the subsequent formation of the Rad51 nucleoprotein filament at DSBs are abolished in Ubc13-deficient cells. Furthermore, generation of ssDNA/RPA complexes at DSBs is severely attenuated in the absence of Ubc13. These data reveal a critical and unexpected role for vertebrate Ubc13 in the initiation of HR at the level of DSB processing.

KW - DNA

KW - PROTEINS

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A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination

Abstract

Keywords

ASJC Scopus subject areas

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