泛素连接酶Brl2在DNA双链断裂修复中的作用分析
收稿日期: 2022-02-14
修回日期: 2022-04-25
网络出版日期: 2022-05-19
基金资助
山西省高等学校科技创新项目编号(2020L0497);山西大同大学博士科研启动经费资助编号(2019-B-20)
The functional analysis of the ubiquitin ligase Brl2 in the repair of DNA double-strand breaks
Received date: 2022-02-14
Revised date: 2022-04-25
Online published: 2022-05-19
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)
组蛋白H2B单泛素化在基因转录、DNA复制及损伤修复中发挥着重要的调控作用。在裂殖酵母(Schizosaccharomyces pombe)中,Brl2作为一个泛素化连接酶,调节H2B的119位赖氨酸的单泛素化。目前,有关Brl2在DNA损伤修复中的作用研究较少,本研究利用药物喜树碱(camptothecin, CPT)处理裂殖酵母产生高毒性的DNA双链断裂(DNA double-strand breaks, DSBs),探索Brl2在DSB修复过程中的作用。研究发现,brl2基因缺失的菌株对CPT高度敏感,并导致细胞内DNA自发重组频率下降。荧光分析表明Brl2和重组修复蛋白Rad52共定位到DSB处,且Brl2促进Rad52在DSB处的募集。在CPT产生的DSB条件下,Brl2会发生磷酸化。以上研究发现揭示了Brl2在DSB修复过程中起重要作用,为具体阐明Brl2在DNA同源重组及双链断裂修复的分子机制奠定了进一步研究的基础。
刘晓琴, 常斐然, 刘思杰, 伍斐, 孔道春 . 泛素连接酶Brl2在DNA双链断裂修复中的作用分析[J]. 遗传, 2022 , 44(7) : 609 -617 . DOI: 10.16288/j.yczz.22-031
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
| [1] | Mehta A, Haber JE. Sources of DNA double-strand breaks and models of recombinational DNA repair. Cold Spring Harb Perspect Biol, 2014, 6(9):a016428. |
| [2] | Huang M, Yang YR, Sun XY, Zhang T, Guo CX. RAD51 regulates REV1 recruitment to DNA double-strand break sites. Hereditas(Beijing), 2018, 40(11):1007-1014. |
| [2] | 黄敏, 杨业然, 孙晓艳, 张婷, 郭彩霞. RAD51调控REV1参与DNA双链断裂修复. 遗传, 2018, 40(11):1007-1014. |
| [3] | Thadathil N, Hori R, Xiao JF, Khan MM. DNA double- strand breaks: a potential therapeutic target for neurodegenerative diseases. Chromosome Res, 2019, 27(4):345-364. |
| [4] | Dai YX, Zhang F, Wang LG, Shan S, Gong ZH, Zhou Z. Structural basis for shieldin complex subunit 3-mediated recruitment of the checkpoint protein REV7 during DNA double-strand break repair. J Biol Chem, 2020, 295(1):250-262. |
| [5] | Shrivastav M, De Haro LP, Nickoloff JA. Regulation of DNA double-strand break repair pathway choice. Cell Res, 2008, 18(1):134-147. |
| [6] | Gong Y, Handa N, Kowalczykowski SC, de Lange T. PHF11 promotes DSB resection, ATR signaling, and HR. Genes Dev, 2017, 31(1):46-58. |
| [7] | Whelan DR, Lee WTC, Marks F, Kong YT, Yin YD, Rothenberg E. Super-resolution visualization of distinct stalled and broken replication fork structures. PLoS Genet, 2020, 16(12):e1009256. |
| [8] | Liu SJ, Hua Y, Wang JN, Li LY, Yuan JJ, Zhang B, Wang ZY, Ji JG, Kong DC. RNA polymerase III is required for the repair of DNA double-strand breaks by homologous recombination. Cell, 2021, 184(5): 1314-1329.e1310. |
| [9] | Limbo O, Porter-Goff ME, Rhind N, Russell P. Mre11 nuclease activity and Ctp1 regulate Chk1 activation by Rad3ATR and Tel1ATM checkpoint kinases at double-strand breaks. Mol Cell Biol, 2011, 31(3):573-583. |
| [10] | Sato H, Niimi A, Yasuhara T, Permata TBM, Hagiwara Y, Isono M, Nuryadi E, Sekine R, Oike T, Kakoti S, Yoshimoto Y, Held KD, Suzuki Y, Kono K, Miyagawa K, Nakano T, Shibata A. DNA double-strand break repair pathway regulates PD-L1 expression in cancer cells. Nat Commun, 2017, 8(1):1751. |
| [11] | Zofall M, Grewal SI. HULC, a histone H2B ubiquitinating complex, modulates heterochromatin independent of histone methylation in fission yeast. J Biol Chem, 2007, 282(19):14065-14072. |
| [12] | Deng ZH, Ai HS, Lu CP, Li JB. The Bre1/Rad6 machinery: writing the central histone ubiquitin mark on H2B and beyond. Chromosome Res, 2020, 28(3-4):247-258. |
| [13] | So CC, Ramachandran S, Martin A. E3 ubiquitin ligases RNF20 and RNF40 are required for double-stranded break (DSB) repair: evidence for monoubiquitination of histone H2B lysine 120 as a novel axis of DSB signaling and repair. Mol Cell Biol, 2019, 39(8):e00488-e00518. |
| [14] | In S, Kim YI, Lee JE, Kim J. RNF20/40-mediated eEF1BδL monoubiquitylation stimulates transcription of heat shock-responsive genes. Nucleic Acids Res, 2019, 47(6):2840-2855. |
| [15] | Scherer S, Davis RW. Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci USA, 1979, 76(10):4951-4955. |
| [16] | Rai SK, Atwood-Moore A, Levin HL. High-frequency lithium acetate transformation of Schizosaccharomyces pombe. Methods Mol Biol, 2018, 1721:167-177. |
| [17] | Forsburg SL, Rhind N. Basic methods for fission yeast. Yeast, 2006, 23(3):173-183. |
| [18] | Osman F, Fortunato EA, Subramani S. Double-strand break-induced mitotic intrachromosomal recombination in the fission yeast Schizosaccharomyces pombe. Genetics, 1996, 142(2):341-357. |
| [19] | Nagy Z, Comer S, Smolenski A. Analysis of protein phosphorylation using Phos-tag gels. Curr Protoc Protein Sci, 2018, 93(1):e64. |
| [20] | Moyal L, Lerenthal Y, Gana-Weisz M, Mass G, So S, Wang SY, Eppink B, Chung YM, Shalev G, Shema E, Shkedy D, Smorodinsky NI, van Vliet N, Kuster B, Mann M, Ciechanover A, Dahm-Daphi J, Kanaar R, Hu MC, Chen DJ, Oren M, Shiloh Y. Requirement of ATM-dependent monoubiquitylation of histone H2B for timely repair of DNA double-strand breaks. Mol Cell, 2011, 41(5):529-542. |
| [21] | Nakamura K, Kato A, Kobayashi J, Yanagihara H, Sakamoto S, Oliveira DVNP, Shimada M, Tauchi H, Suzuki H, Tashiro S, Zou L, Komatsu K. Regulation of homologous recombination by RNF20-dependent H2B ubiquitination. Mol Cell, 2011, 41(5):515-528. |
| [22] | Zheng SH, Li D, Lu Z, Liu GX, Wang M, Xing PY, Wang M, Dong Y, Wang XJ, Li JY, Zhang SM, Peng HY, Ira G, Li GH, Chen XF. Bre1-dependent H2B ubiquitination promotes homologous recombination by stimulating histone eviction at DNA breaks. Nucleic Acids Res, 2018, 46(21):11326-11339. |
| [23] | Zukowski A, Johnson AM. The interplay of histone H2B ubiquitination with budding and fission yeast heterochromatin. Curr Genet, 2018, 64(4):799-806. |
| [24] | Marsh DJ, Ma Y, Dickson KA. Histone monoubiquitination in chromatin remodelling: focus on the histone H2B interactome and cancer. Cancers (Basel), 2020, 12(11):3462. |
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:
