泛素连接酶Brl2在DNA双链断裂修复中的作用分析

doi:10.16288/j.yczz.22-031

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Hereditas(Beijing) ›› 2022, Vol. 44 ›› Issue (7): 609-617.doi: 10.16288/j.yczz.22-031

• Research Article • Previous Articles Next Articles

The functional analysis of the ubiquitin ligase Brl2 in the repair of DNA double-strand breaks

Xiaoqin Liu¹(), Feiran Chang², Sijie Liu², Fei Wu², Daochun Kong²

1. Institute of Brain Science, Shanxi Datong University, Datong 037009, China
2. College of Life Sciences, Peking University, Beijing 100871, China

Received:2022-02-14 Revised:2022-04-25 Online:2022-07-20 Published:2022-05-19
Contact: Liu Xiaoqin E-mail:xiaoqinliu@sxdtdx.edu.cn
Supported by:
Supported by the Scientific and Technological Innovation Program of Higher Education Institutions in Shanxi No(2020L0497);Doctoral Research Start-up Funds of Shanxi Datong University No(2019-B-20)

Abstract

Abstract:

Mono-ubiquitination of histone H2B plays a critical role in the regulation of gene transcription, DNA replication, and DNA damage repair. In Schizosaccharomyces pombe, Brl2 is an E3 ubiquitin ligase and required for the ubiquitination of H2B at lysine residue 119. Currently, there are few studies related to the function of Brl2 in DNA damage repair. Using camptothecin (CPT) to induce DNA double-strand breaks (DSBs) in S. pombe, we investigated the effect of Brl2 on DSB repair, and found that brl2-null mutants showed greater sensitivity to CPT when compared with wild-type (WT) cells, as well as having a drastically reduced spontaneous recombinant frequency. The fluorescent analysis demonstrated that Brl2 was co-localized with the recombination factor Rad52 at DSBs. Moreover, Brl2 promoted the recruitment of Rad52 to DSBs. Under CPT-induced DSBs, Brl2 was phosphorylated. These findings indicate that Brl2 plays a critical role in DNA homologous recombination and its mediated repair of DSBs.

Key words: histone H2B, Brl2, DSB repair, CPT, S. pombe

Xiaoqin Liu, Feiran Chang, Sijie Liu, Fei Wu, Daochun Kong. The functional analysis of the ubiquitin ligase Brl2 in the repair of DNA double-strand breaks[J]. Hereditas(Beijing), 2022, 44(7): 609-617.

Figures/Tables 6

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菌株	交配型	基因型
LD330	h^-	Ura4-D18
rad3Δ	h^-	leu-32, ura4-D18, ade6-M210, his-D1, rad3::leu2
cds1Δ	h⁺	ade6-704, cds1::kanMX6, leu1-32, ura4-D18
brl2Δ	h^-	brl2::ura4, Ura4-D18
brl2Δ rad3Δ	h^-	brl2::ura4, leu-32, ura4-D18, ade6-M210, his-D1, rad3::leu2
brl2Δ cds1Δ	h⁺	brl2::ura4, ade6-704, cds1::kanMX6, leu1-32, ura4-D18
YKM619	h^-	ade6-M375 int:pUC8/ura4⁺/ade6-469 ura4-D18
brl2Δ-YKM619	h^-	brl2::kanMX6, ade6-M375 int: pUC8/ura4⁺/ade6-469 ura4-D18
rad3Δ-YKM619	h^-	rad3::hph, ade6-M375 int: pUC8/ura4⁺/ade6-469 ura4-D18
rad3Δ brl2Δ-YKM619	h^-	brl2::kanMX6, Rad3::hph, ade6-M375 int: pUC8/ura4⁺/ade6-469 ura4-D18
FY18445	h^-	rad52:EYFP:kanr leu1-32 ura4-D18
brl2Δ- FY18445	h^-	brl2::ura4, rad52:EYFP:kanr leu1-32 ura4-D18
DY1012	h^-	Leu-32 his3-D1 ura4-294 arg3Δ::HOsite-natMX ars::P41nmt-HO(his3+)lys-131::Pdis1-mCherry- LacI(lys1+)erg7ter::lacOrepeat(ura4+)rad52-2XCFP::hphMX
brl2-3HA	h^-	brl2::brl2-3HA-ura4, Ura4-D18

The functional analysis of the ubiquitin ligase Brl2 in the repair of DNA double-strand breaks

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