The key role of CYC2 during meiosis in Tetrahymena thermophila

Abstract

Meiotic recombination is carried out through a specialized pathway for the formation and repair of DNA double-strand breaks (DSBs) made by the Spo11 protein. The present study shed light on the functional role of cyclin, CYC2, in Tetrahymena thermophila which has transcriptionally high expression level during meiosis process. Knocking out the CYC2 gene results in arrest of meiotic conjugation process at 2.5–3.5 h after conjugation initiation, before the meiosis division starts, and in company with the absence of DSBs. To investigate the underlying mechanism of this phenomenon, a complete transcriptome profile was performed between wild-type strain and CYC2 knock-out strain. Functional analysis of RNA-Seq results identifies related differentially expressed genes (DEGs) including SPO11 and these DEGs are enriched in DNA repair/mismatch repair (MMR) terms in homologous recombination (HR), which indicates that CYC2 could play a crucial role in meiosis by regulating SPO11 and participating in HR.

Keywords

cyclin / meiosis / RNA-Seq / Tetrahymena thermophila / homologous recombination

References

[1] Bergerat A, de Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P (1997) An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature 386:414–417

[2] Bindea G, Mlecnik B, Hackl H, Charoentong P, Tosolini M, Kirilovsky A, Fridman WH, Pages F, Trajanoski Z, Galon J (2009) ClueGO: a cytoscape plug-into decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics 25:1091–1093

[3] Boateng KA, Bellani MA, Gregoretti IV, Pratto F, Camerini-Otero RD (2013) Homologous pairing preceding SPO11-mediated doublestrand breaks in Mice. Dev Cell 24:196–205

[4] Cao L, Alani E, Kleckner N (1990) A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell 61:1089–1101

[5] Collins K (2012) Perspectives on the ciliated protozoan Tetrahymena thermophila. Method Cell Biol 109:3–7

[6] Devault A, Gueydon E, Schwob E (2008) Interplay between S-cyclindependent kinase and Dbf4-dependent kinase in controlling DNA replication through phosphorylation of yeast Mcm4 N-terminal domain. Mol Biol Cell 19:2267–2277

[7] Gao S, Xiong J, Zhang CC, Berquist BR, Yang RD, Zhao M, Molascon AJ, Kwiatkowski SY, Yuan DX, Qin ZH (2013) Impaired replication elongation in Tetrahymena mutants deficient in histone H3 Lys 27 monomethylation. Gene Dev 27:1662–1679

[8] Gibson TJ, Thompson JD, Blocker A, Kouzarides T (1994) Evidence for a protein domain superfamily shared by the cyclins, Tfiib and Rb/P107. Nucleic Acids Res 22:946–952

[9] Gorovsky MA, Yao MC, Keevert JB, Pleger GL (1975) Isolation of micro- and macronuclei of Tetrahymena pyriformis. Methods Cell Biol 9:311–327

[10] Hayashi A, Mochizuki K (2015) Targeted gene disruption by ectopic induction of DNA elimination in Tetrahymena. Genetics 201:55–64

[11] Henderson KA, Kee K, Maleki S, Santini PA, Keeney S (2006) Cyclin-dependent kinase directly regulates initiation of meiotic recombination. Cell 125:1321–1332

[12] Howard-Till RA, Lukaszewicz A, Loidl J (2011) The recombinases Rad51 and Dmc1 play distinct roles in DNA break repair and recombination partner choice in the meiosis of Tetrahymena. Plos Genet 7:e1001359

[13] Kauppi L, Jeffreys AJ, Keeney S (2004) Where the crossovers are: recombination distributions in mammals. Nat Rev Genet 5:413–424

[14] Keeney S (2001) Mechanism and control of meiotic recombination initiation. Curr Top Dev Biol 52:1–53

[15] Keeney S, Giroux CN, Kleckner N (1997a) Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88:375–384

[16] Keeney S, Giroux CN, Kleckner N (1997b) Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88:375–384

[17] Lam I, Keeney S (2015) Mechanism and regulation of meiotic recombination initiation. Cold Spring Harb Perspect Biol 7: a016634

[18] Loidl J (2013) The hidden talents of SPO11. Dev Cell 24:123–124

[19] Loidl J, Mochizuki K (2009) Tetrahymena meiotic nuclear reorganization is induced by a checkpoint kinase-dependent response to DNA damage. Mol Biol Cell 20:2428–2437

[20] Miao W, Xiong J, Bowen J, Wang W, Liu YF, Braguinets O, Grigull J, Pearlman RE, Orias E, Gorovsky MA (2009) microarray analyses of gene expression during the Tetrahymena thermophila life cycle. Plos One 4:e4429

[21] Mochizuki K, Gorovsky MA (2004) Small RNAs in genome rearrangement in Tetrahymena. Curr Opin Genet Dev 14:181–187

[22] Mochizuki K, Novatchkova M, Loidl J (2008) DNA double-strand breaks, but not crossovers, are required for the reorganization of meiotic nuclei in Tetrahymena. J Cell Sci 121:2148–2158

[23] Petronczki M, Siomos MF, Nasmyth K (2003) Un menage a quatre: the molecular biology of chromosome segregation in meiosis. Cell 112:423–440

[24] Shah JC, Clancy MJ (1992) Ime4, a gene that mediates mat and nutritional control of meiosis in Saccharomyces-cerevisiae. Mol Cell Biol 12:1078–1086

[25] Shodhan A, Lukaszewicz A, Novatchkova M, Loidl J (2014) Msh4 and Msh5 function in SC-independent chiasma formation during the streamlined meiosis of Tetrahymena. Genetics 198:983–993

[26] Song XY, Gjoneska E, Ren QH, Taverna SD, Allis CD, Gorovsky MA (2007) Phosphorylation of the SQ H2A.X motif is required for proper meiosis and mitosis in Tetrahymena thermophila. Mol Cell Biol 27:2648–2660

[27] Spies M, Fishel R (2015) Mismatch repair during homologous and homeologous recombination. Cold Spring Harb Perspect Biol 7: a022657

[28] Stover NA, Rice JD (2011) Distinct cyclin genes define each stage of ciliate conjugation. Cell Cycle 10:1699–1701

[29] Sun H, Treco D, Schultes NP, Szostak JW(1989)Double-strand breaks at an initiation site for meiotic gene conversion. Nature 338:87–90

[30] Szostak JW, Orrweaver TL, Rothstein RJ, Stahl FW(1983) The doublestrand-break repair model for recombination. Cell 33:25–35

[31] Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63

[32] Woehrer SL, Aronica L, Suhren JH, Busch CJL, Noto T, Mochizuki K (2015) A Tetrahymena Hsp90 co-chaperone promotes siRNA loading by ATP-dependent and ATP-independent mechanisms. EMBO J 34:559–577

[33] Xiong J, Lu XY, Zhou ZM, Chang Y, Yuan DX, Tian M, Zhou ZG, Wang L, Fu CJ, Orias E (2012) Transcriptome analysis of the model protozoan, Tetrahymena thermophila, using deep RNA sequencing. Plos One 7:e30630

[34] Zhang H, Adl SM, Berger JD (1999) Two distinct classes of mitotic cyclin homologues, Cyc1 and Cyc2, are involved in cell cycle regulation in the ciliate Paramecium tetraurelia. J Eukaryot Microbiol 46:585–596

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2014 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.