Abstract
In fission yeast, meiotic prophase nuclei develop structures known as linear elements (LinEs), instead of a canonical synaptonemal complex. LinEs contain Rec10 protein. While Rec10 is essential for meiotic recombination, the precise role of LinEs in this process is unknown. Using in situ immunostaining, we show that Rec7 (which is required for meiosis-specific DNA double-strand break (DSB) formation) aggregates in foci on LinEs. The strand exchange protein Rad51, which is known to mark the sites of DSBs, also localizes to LinEs, although to a lesser degree. The number of Rec7 foci corresponds well with the average number of genetic recombination events per meiosis suggesting that Rec7 marks the sites of recombination. Rec7 and Rad51 foci do not co-localize, presumably because they act sequentially on recombination sites. The localization of Rec7 is dependent on Rec10 but independent of the DSB-inducing protein Rec12/Spo11. Neither Rec7 nor Rad51 localization depends on the LinE-associated proteins Hop1 and Mek1, but the formation of Rad51 foci depends on Rec10, Rec7, and, as expected, Rec12/Spo11. We propose that LinEs form around designated recombination sites before the induction of DSBs and that most, if not all, meiotic recombination initiates within the setting provided by LinEs.
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References
Agarwal S, Roeder GS (2000) Zip3 provides a link between recombination enzymes and synaptonemal complex proteins. Cell 102:245–255
Alpi A, Pasierbek P, Gartner A, Loidl J (2003) Genetic and cytological characterization of the recombination protein RAD-51 in Caenorhabditis elegans. Chromosoma 112:6–16
Anderson LK, Royer SM, Page SL, McKim KS, Lai A, Lilly MA, Hawley RS (2005) Juxtaposition of C(2)M and the transverse filament protein C(3)G within the central region of Drosophila synaptonemal complex. Proc Natl Acad Sci U S A 102:4482–4487
Arora C, Kee K, Maleki S, Keeney S (2004) Antiviral protein Ski8 is a direct partner of Spo11 in meiotic DNA break formation, independent of its cytoplasmic role in RNA metabolism. Mol Cell 13:549–559
Aylon Y, Kupiec M (2004) New insights into the mechanism of homologous recombination in yeast. Mutat Res Rev Mutat Res 566:231–248
Bähler J, Wyler T, Loidl J, Kohli J (1993) Unusual nuclear structures in meiotic prophase of fission yeast: a cytological analysis. J Cell Biol 121:241–256
Blat Y, Protacio RU, Hunter N, Kleckner N (2002) Physical and functional interactions among basic chromosome organizational features govern early steps of meiotic chiasma formation. Cell 111:1–20
Bogdanov YF, Dadashev SY, Grishaeva TM (2002) Comparative genomics and proteomics of Drosophila, Brenner’s nematode, and Arabidopsis: identification of functionally similar genes and proteins of meiotic chromosome synapsis. Russ J Genet 38:908–917
Börner GV, Kleckner N, Hunter N (2004) Crossover/noncrossover differentiation, synaptonemal complex formation, and regulatory surveillance at the leptotene/zygotene transition of meiosis. Cell 117:29–45
Cromie GA, Rubio CA, Hyppa RW, Smith GR (2005) A natural meiotic DNA break site in Schizosaccharomyces pombe is a hotspot of gene conversion, highly associated with crossing over. Genetics 169:595–605
Davis L, Smith GR (2001) Meiotic recombination and chromosome segregation in Schizosaccharomyces pombe. Proc Natl Acad Sci U S A 98:8395–8402
Davis L, Barbera M, McDonnell A, McIntyre K, Sternglanz R, Jin Q, Loidl J, Engebrecht J (2001) The Saccharomyces cerevisiae MUM2 gene interacts with the DNA replication machinery and is required for meiotic levels of double strand breaks. Genetics 157:1179–1189
de Massy B (2003) Distribution of meiotic recombination sites. Trends Genet 19:514–522
De Veaux LC, Smith GR (1994) Region-specific activators of meiotic recombination in Schizosaccharomyces pombe. Genes Dev 8:203–210
Ellermeier C, Schmidt H, Smith GR (2004) Swi5 acts in meiotic DNA joint molecule formation in Schizosaccharomyces pombe. Genetics 168:1891–1898
Ellermeier C, Smith GR (2005) Cohesins are required for meiotic DNA breakage and recombination in Schizosaccharomyces pombe. Proc Natl Acad Sci U S A 102:10952–10957
Gregan J, Rabitsch PK, Sakem B, Csutak O, Latypov V, Lehmann E, Kohli J, Nasmyth K (2005) Novel genes required for meiotic chromosome segregation are identified by a high-throughput knockout screen in fission yeast. Curr Biol 15:1663–1669
Grishchuk AL, Kraehenbuehl R, Molnar M, Fleck O, Kohli J (2004) Genetic and cytological characterization of the RecA-homologous proteins Rad51 and Dmc1 of Schizosaccharomyces pombe. Curr Genet 44:317–328
Henderson KA, Keeney S (2004) Tying synaptonemal complex initiation to the formation and programmed repair of DNA double-strand breaks. Proc Natl Acad Sci U S A 101:4519–4524
Kee K, Protacio RU, Arora C, Keeney S (2004) Spatial organization and dynamics of the association of Rec102 and Rec104 with meiotic chromosomes. EMBO J 23:1815–1824
Keeney S (2001) Mechanism and control of meiotic recombination initiation. Curr Top Dev Biol 52:1–53
Kleckner N (1996) Meiosis: how could it work? Proc Natl Acad Sci U S A 93:8167–8174
Kon N, Krawchuk MD, Warren BG, Smith GR, Wahls WP (1997) Transcription factor Mts1/Mts2 (Atf1/Pcr1, Gad7/Pcr1) activates the M26 meiotic recombination hotspot in Schizosaccharomyces pombe. Proc Natl Acad Sci U S A 94:13765–13770
Krawchuk MD, De Veaux LC, Wahls WP (1999) Meiotic chromosome dynamics dependent upon the rec8 +, rec10 + and rec11 + genes of the fission yeast Schizosaccharomyces pombe. Genetics 153:57–68
Lorenz A, Wells JL, Pryce DW, Novatchkova M, Eisenhaber F, McFarlane RJ, Loidl J (2004) S. pombe meiotic linear elements contain proteins related to synaptonemal complex components. J Cell Sci 117:3343–3351
Malone RE, Pittman DL, Nau JJ (1997) Examination of the intron in the meiosis-specific recombination gene REC114 in Saccharomyces. Mol Gen Genet 255:410–419
Miyazaki T, Bressan DA, Shinohara M, Haber JE, Shinohara A (2004) In vivo assembly and disassembly of Rad51 and Rad52 complexes during double-strand break repair. EMBO J 23:939–949
Mizuno K, Emura Y, Baur M, Kohli J, Ohta K, Shibata T (1997) The meiotic recombination hot spot created by the single- base substitution ade6-M26 results in remodeling of chromatin structure in fission yeast. Genes Dev 11:876–886
Molnar M, Parisi S, Kakihara Y, Nojima H, Yamamoto A, Hiraoka Y, Bozsik A, Sipiczki M, Kohli J (2001) Characterization of rec7, an early meiotic recombination gene in Schizosaccharomyces pombe. Genetics 157:519–532
Molnar M, Doll E, Yamamoto A, Hiraoka Y, Kohli J (2003) Linear element formation and their role in meiotic sister chromatid cohesion and chromosome pairing. J Cell Sci 116:1719–1731
Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823
Munz P (1994) An analysis of interference in the fission yeast Schizosaccharomyces pombe. Genetics 137:701–707
Murakami H, Borde V, Shibata T, Lichten M, Ohta K (2003) Correlation between premeiotic DNA replication and chromatin transition at yeast recombination initiation sites. Nucleic Acids Res 31:4085–4090
Neale MJ, Pan J, Keeney S (2005) Endonucleolytic processing of covalent protein-linked DNA double-strand breaks. Nature 436:1053–1057
Öllinger R, Alsheimer M, Benavente R (2005) Mammalian protein SCP1 forms synaptonemal complex-like structures in the absence of meiotic chromosomes. Mol Biol Cell 16:212–217
Olson LW, Edén U, Mitani ME, Egel R (1978) Asynaptic meiosis in fission yeast? Hereditas 89:189–199
Page SL, Hawley RS (2004) The genetics and molecular biology of the synaptonemal complex. Annu Rev Cell Dev Biol 20:525–558
Pâques F, Haber JE (1999) Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 63:349–404
Parisi S, McKay MJ, Molnar M, Thompson MA, van der Spek PJ, van Drunen-Schoenmaker E, Kanaar R, Lehmann E, Hoeijmakers JHJ, Kohli J (1999) Rec8p, a meiotic recombination and sister chromatid cohesion phosphoprotein of the Rad21p family conserved from fission yeast to humans. Mol Cell Biol 19:3515–3528
Pérez-Hidalgo L, Moreno S, San-Segundo PA (2003) Regulation of meiotic progression by the meiosis-specific checkpoint kinase Mek1 in fission yeast. J Cell Sci 116:259–271
Petes TD (2001) Meiotic recombination hot spots and cold spots. Nat Rev Genet 2:360–368
Prieler S, Penkner A, Borde V, Klein F (2005) The control of Spo11’s interaction with meiotic recombination hotspots. Genes Dev 19:255–269
Pryce DW, Lorenz A, Smirnova JB, Loidl J, McFarlane RJ (2005) Differential activation of M26-containing meiotic recombination hot spots in Schizosaccharomyces pombe. Genetics 170:95–106
Reddy KC, Villeneuve AM (2004) C. elegans HIM-17 links chromatin modification and competence for initiation of meiotic recombination. Cell 118:439–452
Roeder GS (1997) Meiotic chromosomes: it takes two to tango. Genes Dev 11:2600–2621
Sauvageau S, Stasiak AZ, Banville I, Ploquin M, Stasiak A, Masson J-Y (2005) Fission yeast Rad51 and Dmc1, two efficient DNA recombinases forming helical nucleoprotein filaments. Mol Cell Biol 25:4377–4387
Scherthan H, Bähler J, Kohli J (1994) Dynamics of chromosome organization and pairing during meiotic prophase in fission yeast. J Cell Biol 127:273–285
Shinohara A, Ogawa H, Ogawa T (1992) Rad51 protein involved in repair and recombination in Saccharomyces cerevisiae is a RecA-like protein. Cell 69:457–470
Thompson DA, Stahl FW (1999) Genetic control of recombination partner preference in yeast meiosis: isolation and characterization of mutants elevated for meiotic unequal sister-chromatid recombination. Genetics 153:621–641
Wan L, de los Santos T, Zhang C, Shokat K, Hollingsworth NM (2004) Mek1 kinase activity functions downstream of RED1 in the regulation of meiotic DSB repair in budding yeast. Mol Biol Cell 15:11–23
Wu T-C, Lichten M (1994) Meiosis-induced double-strand break sites determined by yeast chromatin structure. Science 263:515–517
Yamada T, Mizuno K, Hirota K, Kon N, Wahls WP, Hartsuiker E, Murofushi H, Shibata T, Ohta K (2004) Roles of histone acetylation and chromatin remodeling factor in a meiotic recombination hotspot. EMBO J 23:1792–1803
Young JA, Hyppa RW, Smith GR (2004) Conserved and nonconserved proteins for meiotic DNA breakage and repair in yeasts. Genetics 167:593–605
Zickler D, Kleckner N (1999) Meiotic chromosomes: integrating structure and function. Annu Rev Genet 33:603–754
Acknowledgements
Rec10 antibody was kindly provided by Ramsay McFarlane (Bangor, UK). We thank Eveline Doll (Berne, Switzerland), Pedro San-Segundo (Salamanca, Spain), and Gerald R. Smith (Seattle, WA, USA) for strains. The valuable comments of Ramsay McFarlane and Gerald R. Smith on the manuscript are gratefully acknowledged. We also wish to thank Mario Spirek for help with the Western blotting experiment. This work was supported by the Austrian Science Fund (Grant P18186).
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Lorenz, A., Estreicher, A., Kohli, J. et al. Meiotic recombination proteins localize to linear elements in Schizosaccharomyces pombe . Chromosoma 115, 330–340 (2006). https://doi.org/10.1007/s00412-006-0053-9
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DOI: https://doi.org/10.1007/s00412-006-0053-9