Abstract
Meiosis is a specialized cell division process by which haploid gametes are produced from a diploid mother cell. Reductional chromosome segregation during meiosis I (MI) is achieved by two unique and conserved events: centromeric cohesin protection (CCP) and sister kinetochore mono-orientation (SKM). In Saccharomyces cerevisiae, a meiosis-specific protein Spo13 plays a role in both these centromere-specific events. Despite genome-wide association of Spo13, we failed to detect its function in global processes such as cohesin loading, cohesion establishment and homologs pairing. While Shugoshin (Sgo1) and protein phosphatase 2A (PP2ARts1) play a central role in CCP, it is not fully understood whether Spo13 functions in the process through a Sgo1- PP2ARts1-dependent or -independent mechanism. To delineate this and to find the relative contribution of each of these proteins in CCP and SKM, we meticulously observed the sister chromatid segregation pattern in the wild type, sgo1Δ, rts1Δ and spo13Δ single mutants and in their respective double mutants. We found that Spo13 protects centromeric cohesin through a Sgo1– PP2ARts1-independent mechanism. To our surprise, we observed a hitherto unknown role of Sgo1 in SKM. Further investigation revealed that Sgo1-mediated recruitment of aurora kinase Ipl1 to the centromere facilitates monopolin loading at the kinetochore during MI. Hence, this study uncovers the role of Sgo1 in SKM and demonstartes how the regulators (Sgo1, PP2ARts1, Spo13) work in a coordinated manner to achieve faithful chromosome segregation during meiosis, the failure of which leads to aneuploidy and birth defects.
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References
Agarwal M, Mehta G, Ghosh SK (2015) Role of Ctf3 and COMA subcomplexes in meiosis: implication in maintaining Cse4 at the centromere and numeric spindle poles. Biochim Biophys Acta 1853:671–684. https://doi.org/10.1016/j.bbamcr.2014.12.032
Attner MA, Miller MP, Ee LS, Elkin SK, Amon A (2013) Polo kinase Cdc5 is a central regulator of meiosis I. Proc Natl Acad Sci USA 110:14278–14283. https://doi.org/10.1073/pnas.1311845110
Bhalla N, Dernberg AF (2008) Prelude to a division. Annu Rec Cell Dev Biol 24:397–424. https://doi.org/10.1146/annurev.cellbio.23.090506.123245
Brar GA, Kiburz BM, Zhang Y, Kim JE, White F, Amon A (2006) Rec8 phosphorylation and recombination promote the step-wise loss of cohesins in meiosis. Nature 441:532–536. https://doi.org/10.1038/nature04794
Carlile TM, Amon A (2008) Meiosis I is established through division-specific translational control of a cyclin Cell 133:280–291. https://doi.org/10.1016/j.cell.2008.02.032
Chelysheva L et al (2005) AtREC8 and AtSCC3 are essential to the monopolar orientation of the kinetochores during meiosis. J Cell Sci 118:4621–4632. https://doi.org/10.1242/jcs.02583
Clyne RK, Katis VL, Jessop L, Benjamin KR, Herskowitz I, Lichten M, Nasmyth K (2003) Polo-like kinase Cdc5 promotes chiasmata formation and cosegregation of sister centromeres at meiosis I. Nat Cell Biol 5:480–485. https://doi.org/10.1038/ncb977
Corbett KD, Harrison SC (2012) Molecular architecture of the yeast monopolin complex. Cell Rep 1:583–589. https://doi.org/10.1016/j.celrep.2012.05.012
Ding DQ, Haraguchi T, Hiraoka Y (2016) A cohesin-based structural platform supporting homologous chromosome pairing in meiosis. Curr Genet 62:499–502. https://doi.org/10.1007/s00294-016-0570-x
George AA, Waleorth NC (2016) Microtubule dynamics decoded by the epigenetic state of centromeric chromatin. Curr Genet 62:691–695. https://doi.org/10.1007/s00294-016-0588-0
Hauf S, Watanabe Y (2004) Kinetochore orientation in mitosis and meiosis Cell 119:317–327. https://doi.org/10.1016/j.cell.2004.10.014
Hauf S, Biswas A, Langegger M, Kawashima SA, Tsukahara T, Watanabe Y (2007) Aurora controls sister kinetochore mono-orientation and homolog bi-orientation in meiosis-I. EMBO J 26:4475–4486. https://doi.org/10.1038/sj.emboj.7601880
Iacovella MG, Daly CN, Kelly JS, Michielsen AJ, Clyne RK (2010) Analysis of Polo-like kinase Cdc5 in the meiosis recombination checkpoint Cell cycle (Georgetown. Tex) 9:1182–1193. https://doi.org/10.4161/cc.9.6.11068
Janke C et al (2004) A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins. more markers promoter substitution cassettes Yeast 21:947–962. https://doi.org/10.1002/yea.1142
Katis VL, Galova M, Rabitsch KP, Gregan J, Nasmyth K (2004a) Maintenance of cohesin at centromeres after meiosis I in budding yeast requires a kinetochore-associated protein related to MEI-S332. Curr Biol CB 14:560–572. https://doi.org/10.1016/j.cub.2004.03.001
Katis VL, Matos J, Mori S, Shirahige K, Zachariae W, Nasmyth K (2004b) Spo13 facilitates monopolin recruitment to kinetochores and regulates maintenance of centromeric cohesion during yeast meiosis. Curr Biol CB 14:2183–2196. https://doi.org/10.1016/j.cub.2004.12.020
Katis VL et al (2010) Rec8 phosphorylation by casein kinase 1 and Cdc7-Dbf4 kinase regulates cohesin cleavage by separase during meiosis. Dev cell 18:397–409. https://doi.org/10.1016/j.devcel.2010.01.014
Kawashima SA, Tsukahara T, Langegger M, Hauf S, Kitajima TS, Watanabe Y (2007) Shugoshin enables tension-generating attachment of kinetochores by loading Aurora to centromeres. Genes Dev 21:420–435. https://doi.org/10.1101/gad.1497307
Kawashima SA, Yamagishi Y, Honda T, Ishiguro K, Watanabe Y (2010) Phosphorylation of H2A by Bub1 prevents chromosomal instability through localizing shugoshin. Sci (New York NY) 327:172–177. https://doi.org/10.1126/science.1180189
Kiburz BM et al (2005) The core centromere and Sgo1 establish a 50-kb cohesin-protected domain around centromeres during meiosis. I. Genes Dev 19:3017–3030. https://doi.org/10.1101/gad.1373005
Kim J et al (2015) Meikin is a conserved regulator of meiosis-I-specific kinetochore function. Nature 517:466–471. https://doi.org/10.1038/nature14097
Kitajima TS, Kawashima SA, Watanabe Y (2004) The conserved kinetochore protein shugoshin protects centromeric cohesion during meiosis. Nature 427:510–517. https://doi.org/10.1038/nature02312
Kitajima TS, Sakuno T, Ishiguro K, Iemura S, Natsume T, Kawashima SA, Watanabe Y (2006) Shugoshin collaborates with protein phosphatase 2A to protect cohesin. Nature 441:46–52. https://doi.org/10.1038/nature04663
Klein F, Mahr P, Galova M, Buonomo SB, Michaelis C, Nairz K, Nasmyth K (1999) A central role for cohesins in sister chromatid cohesion, formation of axial elements, and recombination during yeast meiosis. Cell 98:91–103. https://doi.org/10.1016/s0092-8674(00)80609-1
Lee BH, Amon A, Prinz S (2002) Spo13 regulates cohesin cleavage. Genes Dev 16:1672–1681. https://doi.org/10.1101/gad.989302
Lee BH, Kiburz BM, Amon A (2004) Spo13 maintains centromeric cohesion and kinetochore coorientation during meiosis I. Curr Biol CB 14:2168–2182. https://doi.org/10.1016/j.cub.2004.12.033
Lengronne A et al (2004) Cohesin relocation from sites of chromosomal loading to places of convergent transcription. Nature 430:573–578. https://doi.org/10.1038/nature02742
Lin SJ, O’Connell MJ (2017) DNA Topoisomerase II modulates acetyl-regulation of cohesin-mediated chromosome dynamics. Curr Genet 63:923–930. https://doi.org/10.1007/s00294-017-0691-x
Longtine MS et al. (1998) Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14:953–961. https://doi.org/10.1002/(sici)1097-0061(199807)14:10<953::aid-yea293>3.0.co;2-u
Marston AL, Tham WH, Shah H, Amon A (2004) A genome-wide screen identifies genes required for centromeric cohesion. Sci New York NY 303:1367–1370. https://doi.org/10.1126/science.1094220
Matos J et al (2008) Dbf4-dependent CDC7 kinase links DNA replication to the segregation of homologous chromosomes in meiosis. I Cell 135:662–678. https://doi.org/10.1016/j.cell.2008.10.026
McGuinness BE et al (2009) Regulation of APC/C activity in oocytes by a Bub1-dependent spindle assembly checkpoint. Curr Biol CB 19:369–380. https://doi.org/10.1016/j.cub.2009.01.064
Mehta GD, Rizvi SM, Ghosh SK (2012) Cohesin: a guardian of genome integrity. Biochim Biophys Acta 1823:1324–1342. https://doi.org/10.1016/j.bbamcr.2012.05.027
Mehta GD, Agarwal M, Ghosh SK (2014) Functional characterization of kinetochore protein, Ctf19 in meiosis I: an implication of differential impact of Ctf19 on the assembly of mitotic and meiotic kinetochores in Saccharomyces cerevisiae. Mol Microbiol 91:1179–1199. https://doi.org/10.1111/mmi.12527
Meyer RE, Chuong HH, Hild M, Hansen CL, Kinter M, Dawson DS (2015) Ipl1/Aurora-B is necessary for kinetochore restructuring in meiosis I in Saccharomyces cerevisiae. Mol Biol Cell 26:2986–3000. https://doi.org/10.1091/mbc.E15-01-0032
Michaelis C, Ciosk R, Nasmyth K (1997) Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell 91:35–45
Monje-Casas F, Prabhu VR, Lee BH, Boselli M, Amon A (2007) Kinetochore orientation during meiosis is controlled by Aurora B and the monopolin complex. Cell 128:477–490. https://doi.org/10.1016/j.cell.2006.12.040
Ng TM, Waples WG, Lavoie BD, Biggins S (2009) Pericentromeric sister chromatid cohesion promotes kinetochore biorientation. Mol Biol Cell 20:3818–3827. https://doi.org/10.1091/mbc.E09-04-0330
Ng TM et al (2013) Kinetochore function and chromosome segregation rely on critical residues in histones H3 and H4 in budding yeast. Genetics 195:795–807. https://doi.org/10.1534/genetics.113.152082
Peplowska K, Wallek AU, Storchova Z (2014) Sgo1 regulates both condensin and Ipl1/Aurora B to promote chromosome biorientation. PLoS Gene 10:e1004411. https://doi.org/10.1371/journal.pgen.1004411
Pinsky BA, Biggins S (2005) The spindle checkpoint: tension versus attachment. Trends Cell Biol 15:486–493. https://doi.org/10.1016/j.tcb.2005.07.005
Rabitsch KP et al (2003) Kinetochore recruitment of two nucleolar proteins is required for homolog segregation in meiosis. I Dev Cell 4:535–548
Riedel CG et al (2006) Protein phosphatase 2A protects centromeric sister chromatid cohesion during meiosis. I Nature 441:53–61. https://doi.org/10.1038/nature04664
Shonn MA, McCarroll R, Murray AW (2002) Spo13 protects meiotic cohesin at centromeres in meiosis. I Gene Dev 16:1659–1671. https://doi.org/10.1101/gad.975802
Sourirajan A, Lichten M (2008) Polo-like kinase Cdc5 drives exit from pachytene during budding yeast meiosis. Gene Dev 22:2627–2632. https://doi.org/10.1101/gad.1711408
Sullivan M, Morgan DO (2007) A novel destruction sequence targets the meiotic regulator Spo13 for anaphase-promoting complex-dependent degradation in anaphase. I J Biol Chem 282:19710–19715. https://doi.org/10.1074/jbc.M701507200
Sym M, Roeder GS (1995) Zip1-induced changes in synaptonemal complex structure and polycomplex assembly. J Cell Biol 128:455–466
Toth A, Ciosk R, Uhlmann F, Galova M, Schleiffer A, Nasmyth K (1999) Yeast cohesin complex requires a conserved protein, Eco1p(Ctf7), to establish cohesion between sister chromatids during. DNA Replicat Gene Dev 13:320–333
Toth A, Rabitsch KP, Galova M, Schleiffer A, Buonomo SB, Nasmyth K (2000) Functional genomics identifies monopolin: a kinetochore protein required for segregation of homologs during meiosis I Cell 103:1155–1168
Vanoosthuyse V, Prykhozhij S, Hardwick KG (2007) Shugoshin 2 regulates localization of the chromosomal passenger proteins in fission yeast mitosis Mol Biol Cell 18:1657–1669. https://doi.org/10.1091/mbc.E06-10-0890
Watanabe Y (2005) Shugoshin: guardian spirit at the centromere. Curr Opinion Cell Biol 17:590–595. https://doi.org/10.1016/j.ceb.2005.10.003
Xu Z, Cetin B, Anger M, Cho US, Helmhart W, Nasmyth K, Xu W (2009) Structure and function of the PP2A-shugoshin interaction. Mol Cell 35:426–441. https://doi.org/10.1016/j.molcel.2009.06.031
Yokobayashi S, Watanabe Y (2005) The kinetochore protein Moa1 enables cohesion-mediated monopolar attachment at meiosis I. Cell 123:803–817. https://doi.org/10.1016/j.cell.2005.09.013
Yu HG, Koshland D (2007) The Aurora kinase Ipl1 maintains the centromeric localization of PP2A to protect cohesin during meiosis. J Cell Biol 176:911–918. https://doi.org/10.1083/jcb.200609153
Acknowledgements
SKG lab is supported by the Department of Biotechnology, the Department of Science and Technology and the Board of Research in Nuclear Sciences of Govt. of India (Grant Nos. BT/PR13962/BRB/10/798/2010, SR/SO/BB-57/2009 and 37(1)/14/30/2015/BRNS, respectively). GM was supported by a CSIR fellowship (20 − 6/2009(i)EU-IV/329667). GKA was supported by a DST Inspire fellowship (DST/INSPIRE Fellowship/2015/IF150117). APB was supported by SERB-national postdoctoral fellowship (DST No: PDF/2016/000937). We would like to thank Khushboo Sinha for the strain construction.
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Communicated by M. Kupiec.
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Mehta, G., Anbalagan, G.K., Bharati, A.P. et al. An interplay between Shugoshin and Spo13 for centromeric cohesin protection and sister kinetochore mono-orientation during meiosis I in Saccharomyces cerevisiae. Curr Genet 64, 1141–1152 (2018). https://doi.org/10.1007/s00294-018-0832-x
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DOI: https://doi.org/10.1007/s00294-018-0832-x