The possible function of Flp1 in homologous recombination repair in <em>Saccharomyces cerevisiae</em>

Huong Thi Thu Phung; Hoa Luong Hieu Nguyen; Dung Hoang Nguyen; Huong Thi Thu Phung; Hoa Luong Hieu Nguyen; Dung Hoang Nguyen

doi:10.3934/genet.2018.2.161

AIMS Genetics

2018, Volume 5, Issue 2: 161-176. doi: 10.3934/genet.2018.2.161

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Research article

The possible function of Flp1 in homologous recombination repair in Saccharomyces cerevisiae

NTT Hi-Tech Institute, Nguyen Tat Thanh University, Ho Chi Minh city, Vietnam

Received: 11 October 2017 Accepted: 18 March 2018 Published: 03 April 2018

Saccharomyces cerevisiae Mus81 is a structure-selective endonuclease which constitutes an alternative pathway in parallel with the helicase-topoisomerase Sgs1-Top3-Rmi1 complex to resolve a number of DNA intermediates during DNA replication, repair, and homologous recombination. Previously, it was showed that the N-terminal region of Mus81 was required for its in vivo function in a redundant manner with Sgs1; mus81_Δ120N mutant that lacks the first 120 amino acid residues at the N-terminus exhibited synthetic lethality in combination with the loss of SGS1. In this study, the physiologically important role of the N-terminal region of Mus81 in processing toxic intermediates was further investigated. We examined the cellular defect of sgs1Δmus81_Δ100N cells and observed that although viable, the cells became very sensitive to DNA damaging agents. A single-copy suppressor screening to seek for a factor(s) that could rescue the drug sensitivity of sgs1Δmus81_Δ100N cells was performed and revealed that Flp1, a site-specific recombinase 1 encoded on the 2-micron plasmid was a suppressor. Moreover, Flp1 overexpression could partially suppress the drug sensitivity of mus81Δ cells at 37 °C. Our findings suggest a possible function of Flp1 in coordination with Mus81 and Sgs1 to jointly resolve the branched-DNA structures generated in cells attempting to repair DNA damages.
- Flp1,
- genetic screening,
- homologous recombination repair,
- Mus81,
- Sgs1
Citation: Huong Thi Thu Phung, Hoa Luong Hieu Nguyen, Dung Hoang Nguyen. The possible function of Flp1 in homologous recombination repair in Saccharomyces cerevisiae[J]. AIMS Genetics, 2018, 5(2): 161-176. doi: 10.3934/genet.2018.2.161

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Abstract

Saccharomyces cerevisiae Mus81 is a structure-selective endonuclease which constitutes an alternative pathway in parallel with the helicase-topoisomerase Sgs1-Top3-Rmi1 complex to resolve a number of DNA intermediates during DNA replication, repair, and homologous recombination. Previously, it was showed that the N-terminal region of Mus81 was required for its in vivo function in a redundant manner with Sgs1; mus81_Δ120N mutant that lacks the first 120 amino acid residues at the N-terminus exhibited synthetic lethality in combination with the loss of SGS1. In this study, the physiologically important role of the N-terminal region of Mus81 in processing toxic intermediates was further investigated. We examined the cellular defect of sgs1Δmus81_Δ100N cells and observed that although viable, the cells became very sensitive to DNA damaging agents. A single-copy suppressor screening to seek for a factor(s) that could rescue the drug sensitivity of sgs1Δmus81_Δ100N cells was performed and revealed that Flp1, a site-specific recombinase 1 encoded on the 2-micron plasmid was a suppressor. Moreover, Flp1 overexpression could partially suppress the drug sensitivity of mus81Δ cells at 37 °C. Our findings suggest a possible function of Flp1 in coordination with Mus81 and Sgs1 to jointly resolve the branched-DNA structures generated in cells attempting to repair DNA damages.

References

[1]	Kaliraman V, Mullen JR, Fricke WM, et al. (2001) Functional overlap between Sgs1-Top3 and the Mms4-Mus81 endonuclease. Genes Dev 15: 2730–2740. doi: 10.1101/gad.932201
[2]	Nishino T, Komori K, Ishino Y, et al. (2003) X-ray and biochemical anatomy of an archaeal XPF/Rad1/Mus81 family nuclease: similarity between its endonuclease domain and restriction enzymes. Structure 11: 445–457. doi: 10.1016/S0969-2126(03)00046-7
[3]	Blais V, Gao H, Elwell CA, et al. (2004) RNA interference inhibition of Mus81 reduces mitotic recombination in human cells. Mol Biol Cell 15: 552–562.
[4]	Newman M, Murray-Rust J, Lally J, et al. (2005) Structure of an XPF endonuclease with and without DNA suggests a model for substrate recognition. EMBO J 24: 895–905. doi: 10.1038/sj.emboj.7600581
[5]	Boddy MN, Lopez-Girona A, Shanahan P, et al. (2000) Damage tolerance protein Mus81 associates with the FHA1 domain of checkpoint kinase Cds1. Mol Cell Biol 20: 8758–8766. doi: 10.1128/MCB.20.23.8758-8766.2000
[6]	Interthal H, Heyer WD (2000) MUS81 encodes a novel helix-hairpin-helix protein involved in the response to UV- and methylation-induced DNA damage in Saccharomyces cerevisiae. Mol Gen Genet 263: 812–827. doi: 10.1007/s004380000241
[7]	Abraham J, Lemmers B, Hande MP, et al. (2003) Eme1 is involved in DNA damage processing and maintenance of genomic stability in mammalian cells. EMBO J 22: 6137–6147. doi: 10.1093/emboj/cdg580
[8]	Bastin-Shanower SA, Fricke WM, Mullen JR, et al. (2003) The mechanism of Mus81-Mms4 cleavage site selection distinguishes it from the homologous endonuclease Rad1-Rad10. Mol Cell Biol 23: 3487–3496. doi: 10.1128/MCB.23.10.3487-3496.2003
[9]	Chen XB, Melchionna R, Denis CM, et al. (2001) Human Mus81-associated endonuclease cleaves Holliday junctions in vitro. Mol Cell 8: 1117–1127. doi: 10.1016/S1097-2765(01)00375-6
[10]	Boddy MN, Gaillard PH, McDonald WH, et al. (2001) Mus81-Eme1 are essential components of a Holliday junction resolvase. Cell 107: 537–548. doi: 10.1016/S0092-8674(01)00536-0
[11]	Gaillard PH, Noguchi E, Shanahan P, et al. (2003) The endogenous Mus81-Eme1 complex resolves Holliday junctions by a nick and counternick mechanism. Mol Cell 12: 747–759. doi: 10.1016/S1097-2765(03)00342-3
[12]	Constantinou A, Chen XB, McGowan CH, et al. (2002) Holliday junction resolution in human cells: two junction endonucleases with distinct substrate specificities. EMBO J 21: 5577–5585. doi: 10.1093/emboj/cdf554
[13]	Doe CL, Ahn JS, Dixon J, et al. (2002) Mus81-Eme1 and Rqh1 involvement in processing stalled and collapsed replication forks. J Biol Chem 277: 32753–32759. doi: 10.1074/jbc.M202120200
[14]	Whitby MC, Osman F, Dixon J (2003) Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4. J Biol Chem 278: 6928–6935. doi: 10.1074/jbc.M210006200
[15]	Gaskell LJ, Osman F, Gilbert RJ, et al. (2007) Mus81 cleavage of Holliday junctions: a failsafe for processing meiotic recombination intermediates? EMBO J 26: 1891–1901. doi: 10.1038/sj.emboj.7601645
[16]	Schwartz EK, Wright WD, Ehmsen KT, et al. (2012) Mus81-Mms4 functions as a single heterodimer to cleave nicked intermediates in recombinational DNA repair. Mol Cell Biol 32: 3065–3080. doi: 10.1128/MCB.00547-12
[17]	Matos J, Blanco MG, Maslen S, et al. (2011) Regulatory control of the resolution of DNA recombination intermediates during meiosis and mitosis. Cell 147: 158–172. doi: 10.1016/j.cell.2011.08.032
[18]	Gallo-Fernandez M, Saugar I, Ortiz-Bazan MA, et al. (2012) Cell cycle-dependent regulation of the nuclease activity of Mus81-Eme1/Mms4. Nucleic Acids Res 40: 8325–8335. doi: 10.1093/nar/gks599
[19]	Princz LN, Wild P, Bittmann J, et al. (2017) Dbf4-dependent kinase and the Rtt107 scaffold promote Mus81-Mms4 resolvase activation during mitosis. EMBO J 36: 664–678. doi: 10.15252/embj.201694831
[20]	Saugar I, Jimenez-Martin A, Tercero JA (2017) Subnuclear Relocalization of Structure-Specific Endonucleases in Response to DNA Damage. Cell Rep 20: 1553–1562. doi: 10.1016/j.celrep.2017.07.059
[21]	Mullen JR, Kaliraman V, Ibrahim SS, et al. (2001) Requirement for three novel protein complexes in the absence of the Sgs1 DNA helicase in Saccharomyces cerevisiae. Genetics 157: 103–118.
[22]	Chu WK, Hickson ID (2009) RecQ helicases: multifunctional genome caretakers. Nat Rev Cancer 9: 644–654. doi: 10.1038/nrc2682
[23]	Fabre F, Chan A, Heyer WD, et al. (2002) Alternate pathways involving Sgs1/Top3, Mus81/ Mms4, and Srs2 prevent formation of toxic recombination intermediates from single-stranded gaps created by DNA replication. Proc Natl Acad Sci U S A 99: 16887–16892. doi: 10.1073/pnas.252652399
[24]	Ciccia A, Constantinou A, West SC (2003) Identification and characterization of the human mus81-eme1 endonuclease. J Biol Chem 278: 25172–25178. doi: 10.1074/jbc.M302882200
[25]	Kang MJ, Lee CH, Kang YH, et al. (2010) Genetic and functional interactions between Mus81-Mms4 and Rad27. Nucleic Acids Res 38: 7611–7625. doi: 10.1093/nar/gkq651
[26]	Thu HPT, Nguyen TA, Munashingha PR, et al. (2015) A physiological significance of the functional interaction between Mus81 and Rad27 in homologous recombination repair. Nucleic Acids Res 43: 1684–1699. doi: 10.1093/nar/gkv025
[27]	Chavdarova M, Marini V, Sisakova A, et al. (2015) Srs2 promotes Mus81-Mms4-mediated resolution of recombination intermediates. Nucleic Acids Res 43: 3626–3642. doi: 10.1093/nar/gkv198
[28]	Sisakova A, Altmannova V, Sebesta M, et al. (2017) Role of PCNA and RFC in promoting Mus81-complex activity. BMC Biol 15: 90. doi: 10.1186/s12915-017-0429-8
[29]	Sebesta M, Urulangodi M, Stefanovie B, et al. (2017) Esc2 promotes Mus81 complex-activity via its SUMO-like and DNA binding domains. Nucleic Acids Res 45: 215–230. doi: 10.1093/nar/gkw882
[30]	Keyamura K, Arai K, Hishida T (2016) Srs2 and Mus81-Mms4 Prevent Accumulation of Toxic Inter-Homolog Recombination Intermediates. PLoS Genet 12: e1006136. doi: 10.1371/journal.pgen.1006136
[31]	Ghamrasni SE, Cardoso R, Li L, et al. (2016) Rad54 and Mus81 cooperation promotes DNA damage repair and restrains chromosome missegregation. Oncogene 35: 4836–4845. doi: 10.1038/onc.2016.16
[32]	Ii M, Ii T, Brill SJ (2007) Mus81 functions in the quality control of replication forks at the rDNA and is involved in the maintenance of rDNA repeat number in Saccharomyces cerevisiae. Mutat Res 625: 1–19. doi: 10.1016/j.mrfmmm.2007.04.007
[33]	Futcher AB (1988) The 2 micron circle plasmid of Saccharomyces cerevisiae. Yeast 4: 27–40. doi: 10.1002/yea.320040104
[34]	Chan KM, Liu YT, Ma CH, et al. (2013) The 2 micron plasmid of Saccharomyces cerevisiae: a miniaturized selfish genome with optimized functional competence. Plasmid 70: 2–17. doi: 10.1016/j.plasmid.2013.03.001
[35]	Ahn YT, Wu XL, Biswal S, et al. (1997) The 2microm-plasmid-encoded Rep1 and Rep2 proteins interact with each other and colocalize to the Saccharomyces cerevisiae nucleus. J Bacteriol 179: 7497–7506. doi: 10.1128/jb.179.23.7497-7506.1997
[36]	Velmurugan S, Ahn YT, Yang XM, et al. (1998) The 2 micrometer plasmid stability system: analyses of the interactions among plasmid- and host-encoded components. Mol Cell Biol 18: 7466–7477. doi: 10.1128/MCB.18.12.7466
[37]	Velmurugan S, Yang XM, Chan CS, et al. (2000) Partitioning of the 2-microm circle plasmid of Saccharomyces cerevisiae. Functional coordination with chromosome segregation and plasmid-encoded rep protein distribution. J Cell Biol 149: 553–566.
[38]	Ghosh SK, Hajra S, Paek A, et al. (2006) Mechanisms for chromosome and plasmid segregation. Annu Rev Biochem 75: 211–241. doi: 10.1146/annurev.biochem.75.101304.124037
[39]	Andrews BJ, Proteau GA, Beatty LG, et al. (1985) The FLP recombinase of the 2 micron circle DNA of yeast: interaction with its target sequences. Cell 40: 795–803. doi: 10.1016/0092-8674(85)90339-3
[40]	Gronostajski RM, Sadowski PD (1985) The FLP recombinase of the Saccharomyces cerevisiae 2 microns plasmid attaches covalently to DNA via a phosphotyrosyl linkage. Mol Cell Biol 5: 3274–3279. doi: 10.1128/MCB.5.11.3274

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