Abstract
Spore formation in Saccharomyces cerevisiae requires the de novo synthesis of prospore membranes and spore walls. Ady3p has been identified as an interaction partner for Mpc70p/Spo21p, a meiosis-specific component of the outer plaque of the spindle pole body (SPB) that is required for prospore membrane formation, and for Don1p, which forms a ring-like structure at the leading edge of the prospore membrane during meiosis II. ADY3 expression has been shown to be induced in midsporulation. We report here that Ady3p interacts with additional components of the outer and central plaques of the SPB in the two-hybrid assay. Cells that lack ADY3 display a decrease in sporulation efficiency, and most ady3Delta/ady3Delta asci that do form contain fewer than four spores. The sporulation defect in ady3Delta/ady3Delta cells is due to a failure to synthesize spore wall polymers. Ady3p forms ring-like structures around meiosis II spindles that colocalize with those formed by Don1p, and Don1p rings are absent during meiosis II in ady3Delta/ady3Delta cells. In mpc70Delta/mpc70Delta cells, Ady3p remains associated with SPBs during meiosis II. Our results suggest that Ady3p mediates assembly of the Don1p-containing structure at the leading edge of the prospore membrane via interaction with components of the SPB and that this structure is involved in spore wall formation.
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- Bajgier B. K., Malzone M., Nickas M., Neiman A. M. SPO21 is required for meiosis-specific modification of the spindle pole body in yeast. Mol Biol Cell. 2001 Jun;12(6):1611–1621. doi: 10.1091/mbc.12.6.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Briza P., Breitenbach M., Ellinger A., Segall J. Isolation of two developmentally regulated genes involved in spore wall maturation in Saccharomyces cerevisiae. Genes Dev. 1990 Oct;4(10):1775–1789. doi: 10.1101/gad.4.10.1775. [DOI] [PubMed] [Google Scholar]
- Briza P., Ellinger A., Winkler G., Breitenbach M. Characterization of a DL-dityrosine-containing macromolecule from yeast ascospore walls. J Biol Chem. 1990 Sep 5;265(25):15118–15123. [PubMed] [Google Scholar]
- Briza P., Ellinger A., Winkler G., Breitenbach M. Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan. J Biol Chem. 1988 Aug 15;263(23):11569–11574. [PubMed] [Google Scholar]
- Briza P., Winkler G., Kalchhauser H., Breitenbach M. Dityrosine is a prominent component of the yeast ascospore wall. A proof of its structure. J Biol Chem. 1986 Mar 25;261(9):4288–4294. [PubMed] [Google Scholar]
- Christodoulidou A., Bouriotis V., Thireos G. Two sporulation-specific chitin deacetylase-encoding genes are required for the ascospore wall rigidity of Saccharomyces cerevisiae. J Biol Chem. 1996 Dec 6;271(49):31420–31425. doi: 10.1074/jbc.271.49.31420. [DOI] [PubMed] [Google Scholar]
- Chu S., DeRisi J., Eisen M., Mulholland J., Botstein D., Brown P. O., Herskowitz I. The transcriptional program of sporulation in budding yeast. Science. 1998 Oct 23;282(5389):699–705. doi: 10.1126/science.282.5389.699. [DOI] [PubMed] [Google Scholar]
- Davidow L. S., Goetsch L., Byers B. Preferential Occurrence of Nonsister Spores in Two-Spored Asci of SACCHAROMYCES CEREVISIAE: Evidence for Regulation of Spore-Wall Formation by the Spindle Pole Body. Genetics. 1980 Mar;94(3):581–595. doi: 10.1093/genetics/94.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elliott S., Knop M., Schlenstedt G., Schiebel E. Spc29p is a component of the Spc110p subcomplex and is essential for spindle pole body duplication. Proc Natl Acad Sci U S A. 1999 May 25;96(11):6205–6210. doi: 10.1073/pnas.96.11.6205. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esposito M. S., Esposoto R. E., Moens P. B. Genetic analysis of two spored asci produced by the spo 3 mutant of Saccharomyces. Mol Gen Genet. 1974;135(2):91–95. doi: 10.1007/BF00264777. [DOI] [PubMed] [Google Scholar]
- Gyuris J., Golemis E., Chertkov H., Brent R. Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell. 1993 Nov 19;75(4):791–803. doi: 10.1016/0092-8674(93)90498-f. [DOI] [PubMed] [Google Scholar]
- Inouye C., Dhillon N., Durfee T., Zambryski P. C., Thorner J. Mutational analysis of STE5 in the yeast Saccharomyces cerevisiae: application of a differential interaction trap assay for examining protein-protein interactions. Genetics. 1997 Oct;147(2):479–492. doi: 10.1093/genetics/147.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ishihara S., Hirata A., Minemura M., Nogami S., Ohya Y. A mutation in SPC42, which encodes a component of the spindle pole body, results in production of two-spored asci in Saccharomyces cerevisiae. Mol Genet Genomics. 2001 Jun;265(4):585–595. doi: 10.1007/s004380000442. [DOI] [PubMed] [Google Scholar]
- Ito T., Chiba T., Ozawa R., Yoshida M., Hattori M., Sakaki Y. A comprehensive two-hybrid analysis to explore the yeast protein interactome. Proc Natl Acad Sci U S A. 2001 Mar 13;98(8):4569–4574. doi: 10.1073/pnas.061034498. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klapholz S., Esposito R. E. Isolation of SPO12-1 and SPO13-1 from a natural variant of yeast that undergoes a single meiotic division. Genetics. 1980 Nov;96(3):567–588. doi: 10.1093/genetics/96.3.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knop M., Siegers K., Pereira G., Zachariae W., Winsor B., Nasmyth K., Schiebel E. Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines. Yeast. 1999 Jul;15(10B):963–972. doi: 10.1002/(SICI)1097-0061(199907)15:10B<963::AID-YEA399>3.0.CO;2-W. [DOI] [PubMed] [Google Scholar]
- Knop M., Strasser K. Role of the spindle pole body of yeast in mediating assembly of the prospore membrane during meiosis. EMBO J. 2000 Jul 17;19(14):3657–3667. doi: 10.1093/emboj/19.14.3657. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krisak L., Strich R., Winters R. S., Hall J. P., Mallory M. J., Kreitzer D., Tuan R. S., Winter E. SMK1, a developmentally regulated MAP kinase, is required for spore wall assembly in Saccharomyces cerevisiae. Genes Dev. 1994 Sep 15;8(18):2151–2161. doi: 10.1101/gad.8.18.2151. [DOI] [PubMed] [Google Scholar]
- Longtine M. S., McKenzie A., 3rd, Demarini D. J., Shah N. G., Wach A., Brachat A., Philippsen P., Pringle J. R. Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast. 1998 Jul;14(10):953–961. doi: 10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]
- Moreno-Borchart A. C., Strasser K., Finkbeiner M. G., Shevchenko A., Shevchenko A., Knop M. Prospore membrane formation linked to the leading edge protein (LEP) coat assembly. EMBO J. 2001 Dec 17;20(24):6946–6957. doi: 10.1093/emboj/20.24.6946. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nag D. K., Koonce M. P., Axelrod J. SSP1, a gene necessary for proper completion of meiotic divisions and spore formation in Saccharomyces cerevisiae. Mol Cell Biol. 1997 Dec;17(12):7029–7039. doi: 10.1128/mcb.17.12.7029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neiman A. M., Katz L., Brennwald P. J. Identification of domains required for developmentally regulated SNARE function in Saccharomyces cerevisiae. Genetics. 2000 Aug;155(4):1643–1655. doi: 10.1093/genetics/155.4.1643. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neiman A. M. Prospore membrane formation defines a developmentally regulated branch of the secretory pathway in yeast. J Cell Biol. 1998 Jan 12;140(1):29–37. doi: 10.1083/jcb.140.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pammer M., Briza P., Ellinger A., Schuster T., Stucka R., Feldmann H., Breitenbach M. DIT101 (CSD2, CAL1), a cell cycle-regulated yeast gene required for synthesis of chitin in cell walls and chitosan in spore walls. Yeast. 1992 Dec;8(12):1089–1099. doi: 10.1002/yea.320081211. [DOI] [PubMed] [Google Scholar]
- Primig M., Williams R. M., Winzeler E. A., Tevzadze G. G., Conway A. R., Hwang S. Y., Davis R. W., Esposito R. E. The core meiotic transcriptome in budding yeasts. Nat Genet. 2000 Dec;26(4):415–423. doi: 10.1038/82539. [DOI] [PubMed] [Google Scholar]
- Rabitsch K. P., Tóth A., Gálová M., Schleiffer A., Schaffner G., Aigner E., Rupp C., Penkner A. M., Moreno-Borchart A. C., Primig M. A screen for genes required for meiosis and spore formation based on whole-genome expression. Curr Biol. 2001 Jul 10;11(13):1001–1009. doi: 10.1016/s0960-9822(01)00274-3. [DOI] [PubMed] [Google Scholar]
- Straight P. D., Giddings T. H., Jr, Winey M. Mps1p regulates meiotic spindle pole body duplication in addition to having novel roles during sporulation. Mol Biol Cell. 2000 Oct;11(10):3525–3537. doi: 10.1091/mbc.11.10.3525. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tachikawa H., Bloecher A., Tatchell K., Neiman A. M. A Gip1p-Glc7p phosphatase complex regulates septin organization and spore wall formation. J Cell Biol. 2001 Nov 26;155(5):797–808. doi: 10.1083/jcb.200107008. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tu J., Carlson M. The GLC7 type 1 protein phosphatase is required for glucose repression in Saccharomyces cerevisiae. Mol Cell Biol. 1994 Oct;14(10):6789–6796. doi: 10.1128/mcb.14.10.6789. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tu J., Song W., Carlson M. Protein phosphatase type 1 interacts with proteins required for meiosis and other cellular processes in Saccharomyces cerevisiae. Mol Cell Biol. 1996 Aug;16(8):4199–4206. doi: 10.1128/mcb.16.8.4199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uetz P., Giot L., Cagney G., Mansfield T. A., Judson R. S., Knight J. R., Lockshon D., Narayan V., Srinivasan M., Pochart P. A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature. 2000 Feb 10;403(6770):623–627. doi: 10.1038/35001009. [DOI] [PubMed] [Google Scholar]
- Wesp A., Prinz S., Fink G. R. Conservative duplication of spindle poles during meiosis in Saccharomyces cerevisiae. J Bacteriol. 2001 Apr;183(7):2372–2375. doi: 10.1128/JB.183.7.2372-2375.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]