Abstract
Loss of the nonessential RNA-binding domain protein, Scw1, increases resistance to cell-wall-degrading enzymes in fission yeast. Surprisingly, scw1 null mutations also suppress the lethality of mutations (cdc11-136, cdc7-24, cdc14-118, sid1-239, sid2-250, sid3-106, sid4-A1, and mob1-1) at all levels of the sid pathway. This pathway forms part of the septation initiation network (SIN), which regulates the onset of septum formation and ensures the proper coupling of mitosis to cytokinesis. In contrast, scw1(-) mutations do not suppress ts alleles of the rng genes, cdc12 or cdc15. These mutations also prevent the formation of a septum and in addition block assembly and/or function of the contractile acto-myosin ring. sid mutants exhibit a hyper-sensitivity to cell-wall-degrading enzymes that is suppressed by loss of Scw1. Furthermore, scw1(-)-mediated rescue of sid mutants is abolished in the presence of calcofluor white, a compound that interferes with cell-wall synthesis. These data suggest that Scw1 acts in opposition to the SIN as a negative regulator of cell-wall/septum deposition. Unlike components of the SIN, Scw1 is predominantly a cytoplasmic protein and is not localized to the spindle pole body.
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- Abe H., Shimoda C. Autoregulated expression of Schizosaccharomyces pombe meiosis-specific transcription factor Mei4 and a genome-wide search for its target genes. Genetics. 2000 Apr;154(4):1497–1508. doi: 10.1093/genetics/154.4.1497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Balasubramanian M. K., Helfman D. M., Hemmingsen S. M. A new tropomyosin essential for cytokinesis in the fission yeast S. pombe. Nature. 1992 Nov 5;360(6399):84–87. doi: 10.1038/360084a0. [DOI] [PubMed] [Google Scholar]
- Balasubramanian M. K., McCollum D., Chang L., Wong K. C., Naqvi N. I., He X., Sazer S., Gould K. L. Isolation and characterization of new fission yeast cytokinesis mutants. Genetics. 1998 Jul;149(3):1265–1275. doi: 10.1093/genetics/149.3.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bardin A. J., Amon A. Men and sin: what's the difference? Nat Rev Mol Cell Biol. 2001 Nov;2(11):815–826. doi: 10.1038/35099020. [DOI] [PubMed] [Google Scholar]
- Basi G., Schmid E., Maundrell K. TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. Gene. 1993 Jan 15;123(1):131–136. doi: 10.1016/0378-1119(93)90552-e. [DOI] [PubMed] [Google Scholar]
- Beltraminelli N., Murone M., Simanis V. The S. pombe zfs1 gene is required to prevent septation if mitotic progression is inhibited. J Cell Sci. 1999 Sep;112(Pt 18):3103–3114. doi: 10.1242/jcs.112.18.3103. [DOI] [PubMed] [Google Scholar]
- Bezanilla M., Forsburg S. L., Pollard T. D. Identification of a second myosin-II in Schizosaccharomyces pombe: Myp2p is conditionally required for cytokinesis. Mol Biol Cell. 1997 Dec;8(12):2693–2705. doi: 10.1091/mbc.8.12.2693. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bähler J., Steever A. B., Wheatley S., Wang Y. l., Pringle J. R., Gould K. L., McCollum D. Role of polo kinase and Mid1p in determining the site of cell division in fission yeast. J Cell Biol. 1998 Dec 14;143(6):1603–1616. doi: 10.1083/jcb.143.6.1603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bähler J., Wu J. Q., Longtine M. S., Shah N. G., McKenzie A., 3rd, Steever A. B., Wach A., Philippsen P., Pringle J. R. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast. 1998 Jul;14(10):943–951. doi: 10.1002/(SICI)1097-0061(199807)14:10<943::AID-YEA292>3.0.CO;2-Y. [DOI] [PubMed] [Google Scholar]
- Chang F., Drubin D., Nurse P. cdc12p, a protein required for cytokinesis in fission yeast, is a component of the cell division ring and interacts with profilin. J Cell Biol. 1997 Apr 7;137(1):169–182. doi: 10.1083/jcb.137.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang F., Woollard A., Nurse P. Isolation and characterization of fission yeast mutants defective in the assembly and placement of the contractile actin ring. J Cell Sci. 1996 Jan;109(Pt 1):131–142. doi: 10.1242/jcs.109.1.131. [DOI] [PubMed] [Google Scholar]
- Chang L., Gould K. L. Sid4p is required to localize components of the septation initiation pathway to the spindle pole body in fission yeast. Proc Natl Acad Sci U S A. 2000 May 9;97(10):5249–5254. doi: 10.1073/pnas.97.10.5249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Decottignies A., Zarzov P., Nurse P. In vivo localisation of fission yeast cyclin-dependent kinase cdc2p and cyclin B cdc13p during mitosis and meiosis. J Cell Sci. 2001 Jul;114(Pt 14):2627–2640. doi: 10.1242/jcs.114.14.2627. [DOI] [PubMed] [Google Scholar]
- Edamatsu M., Toyoshima Y. Y. Isolation and characterization of pos mutants defective in correct positioning of septum in Schizosaccharomyces pombe. Zoolog Sci. 1996 Apr;13(2):235–239. doi: 10.2108/zsj.13.235. [DOI] [PubMed] [Google Scholar]
- Evans D. R., Hemmings B. A. Mutation of the C-terminal leucine residue of PP2Ac inhibits PR55/B subunit binding and confers supersensitivity to microtubule destabilization in Saccharomyces cerevisiae. Mol Gen Genet. 2000 Nov;264(4):425–432. doi: 10.1007/s004380000302. [DOI] [PubMed] [Google Scholar]
- Fankhauser C., Marks J., Reymond A., Simanis V. The S. pombe cdc16 gene is required both for maintenance of p34cdc2 kinase activity and regulation of septum formation: a link between mitosis and cytokinesis? EMBO J. 1993 Jul;12(7):2697–2704. doi: 10.1002/j.1460-2075.1993.tb05931.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fankhauser C., Reymond A., Cerutti L., Utzig S., Hofmann K., Simanis V. The S. pombe cdc15 gene is a key element in the reorganization of F-actin at mitosis. Cell. 1995 Aug 11;82(3):435–444. doi: 10.1016/0092-8674(95)90432-8. [DOI] [PubMed] [Google Scholar]
- Fankhauser C., Simanis V. The cdc7 protein kinase is a dosage dependent regulator of septum formation in fission yeast. EMBO J. 1994 Jul 1;13(13):3011–3019. doi: 10.1002/j.1460-2075.1994.tb06600.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feoktistova A., Magnelli P., Abeijon C., Perez P., Lester R. L., Dickson R. C., Gould K. L. Coordination between fission yeast glucan formation and growth requires a sphingolipase activity. Genetics. 2001 Aug;158(4):1397–1411. doi: 10.1093/genetics/158.4.1397. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furge K. A., Cheng Q. C., Jwa M., Shin S., Song K., Albright C. F. Regions of Byr4, a regulator of septation in fission yeast, that bind Spg1 or Cdc16 and form a two-component GTPase-activating protein with Cdc16. J Biol Chem. 1999 Apr 16;274(16):11339–11343. doi: 10.1074/jbc.274.16.11339. [DOI] [PubMed] [Google Scholar]
- Furge K. A., Wong K., Armstrong J., Balasubramanian M., Albright C. F. Byr4 and Cdc16 form a two-component GTPase-activating protein for the Spg1 GTPase that controls septation in fission yeast. Curr Biol. 1998 Aug 27;8(17):947–954. doi: 10.1016/s0960-9822(98)70394-x. [DOI] [PubMed] [Google Scholar]
- Garí E., Volpe T., Wang H., Gallego C., Futcher B., Aldea M. Whi3 binds the mRNA of the G1 cyclin CLN3 to modulate cell fate in budding yeast. Genes Dev. 2001 Nov 1;15(21):2803–2808. doi: 10.1101/gad.203501. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gould K. L., Simanis V. The control of septum formation in fission yeast. Genes Dev. 1997 Nov 15;11(22):2939–2951. doi: 10.1101/gad.11.22.2939. [DOI] [PubMed] [Google Scholar]
- Grallert A., Grallert B., Zilahi E., Szilagyi Z., Sipiczki M. Eleven novel sep genes of Schizosaccharomyces pombe required for efficient cell separation and sexual differentiation. Yeast. 1999 Jun 15;15(8):669–686. doi: 10.1002/(SICI)1097-0061(19990615)15:8<669::AID-YEA411>3.0.CO;2-K. [DOI] [PubMed] [Google Scholar]
- Guertin D. A., Chang L., Irshad F., Gould K. L., McCollum D. The role of the sid1p kinase and cdc14p in regulating the onset of cytokinesis in fission yeast. EMBO J. 2000 Apr 17;19(8):1803–1815. doi: 10.1093/emboj/19.8.1803. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haigler C. H., Brown R. M., Jr, Benziman M. Calcofluor white ST Alters the in vivo assembly of cellulose microfibrils. Science. 1980 Nov 21;210(4472):903–906. doi: 10.1126/science.7434003. [DOI] [PubMed] [Google Scholar]
- Hou M. C., Salek J., McCollum D. Mob1p interacts with the Sid2p kinase and is required for cytokinesis in fission yeast. Curr Biol. 2000 May 18;10(10):619–622. doi: 10.1016/s0960-9822(00)00492-9. [DOI] [PubMed] [Google Scholar]
- Jiang W., Hallberg R. L. Correct regulation of the septation initiation network in Schizosaccharomyces pombe requires the activities of par1 and par2. Genetics. 2001 Aug;158(4):1413–1429. doi: 10.1093/genetics/158.4.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jiménez J., Cid V. J., Cenamor R., Yuste M., Molero G., Nombela C., Sánchez M. Morphogenesis beyond cytokinetic arrest in Saccharomyces cerevisiae. J Cell Biol. 1998 Dec 14;143(6):1617–1634. doi: 10.1083/jcb.143.6.1617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kinoshita K., Nemoto T., Nabeshima K., Kondoh H., Niwa H., Yanagida M. The regulatory subunits of fission yeast protein phosphatase 2A (PP2A) affect cell morphogenesis, cell wall synthesis and cytokinesis. Genes Cells. 1996 Jan;1(1):29–45. doi: 10.1046/j.1365-2443.1996.02002.x. [DOI] [PubMed] [Google Scholar]
- Kippert F., Lloyd D. The aniline blue fluorochrome specifically stains the septum of both live and fixed Schizosaccharomyces pombe cells. FEMS Microbiol Lett. 1995 Oct 15;132(3):215–219. doi: 10.1111/j.1574-6968.1995.tb07836.x. [DOI] [PubMed] [Google Scholar]
- Kitayama C., Sugimoto A., Yamamoto M. Type II myosin heavy chain encoded by the myo2 gene composes the contractile ring during cytokinesis in Schizosaccharomyces pombe. J Cell Biol. 1997 Jun 16;137(6):1309–1319. doi: 10.1083/jcb.137.6.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krapp A., Schmidt S., Cano E., Simanis V. S. pombe cdc11p, together with sid4p, provides an anchor for septation initiation network proteins on the spindle pole body. Curr Biol. 2001 Oct 16;11(20):1559–1568. doi: 10.1016/s0960-9822(01)00478-x. [DOI] [PubMed] [Google Scholar]
- Le Goff X., Buvelot S., Salimova E., Guerry F., Schmidt S., Cueille N., Cano E., Simanis V. The protein phosphatase 2A B'-regulatory subunit par1p is implicated in regulation of the S. pombe septation initiation network. FEBS Lett. 2001 Nov 9;508(1):136–142. doi: 10.1016/s0014-5793(01)03047-2. [DOI] [PubMed] [Google Scholar]
- Le Goff X., Utzig S., Simanis V. Controlling septation in fission yeast: finding the middle, and timing it right. Curr Genet. 1999 Jul;35(6):571–584. doi: 10.1007/s002940050455. [DOI] [PubMed] [Google Scholar]
- Levin D. E., Bishop J. M. A putative protein kinase gene (kin1+) is important for growth polarity in Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8272–8276. doi: 10.1073/pnas.87.21.8272. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu J., Wang H., McCollum D., Balasubramanian M. K. Drc1p/Cps1p, a 1,3-beta-glucan synthase subunit, is essential for division septum assembly in Schizosaccharomyces pombe. Genetics. 1999 Nov;153(3):1193–1203. doi: 10.1093/genetics/153.3.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lussier M., White A. M., Sheraton J., di Paolo T., Treadwell J., Southard S. B., Horenstein C. I., Chen-Weiner J., Ram A. F., Kapteyn J. C. Large scale identification of genes involved in cell surface biosynthesis and architecture in Saccharomyces cerevisiae. Genetics. 1997 Oct;147(2):435–450. doi: 10.1093/genetics/147.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marks J., Fankhauser C., Simanis V. Genetic interactions in the control of septation in Schizosaccharomyces pombe. J Cell Sci. 1992 Apr;101(Pt 4):801–808. doi: 10.1242/jcs.101.4.801. [DOI] [PubMed] [Google Scholar]
- Maundrell K. Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene. 1993 Jan 15;123(1):127–130. doi: 10.1016/0378-1119(93)90551-d. [DOI] [PubMed] [Google Scholar]
- May K. M., Watts F. Z., Jones N., Hyams J. S. Type II myosin involved in cytokinesis in the fission yeast, Schizosaccharomyces pombe. Cell Motil Cytoskeleton. 1997;38(4):385–396. doi: 10.1002/(SICI)1097-0169(1997)38:4<385::AID-CM8>3.0.CO;2-2. [DOI] [PubMed] [Google Scholar]
- McCollum D., Balasubramanian M. K., Pelcher L. E., Hemmingsen S. M., Gould K. L. Schizosaccharomyces pombe cdc4+ gene encodes a novel EF-hand protein essential for cytokinesis. J Cell Biol. 1995 Aug;130(3):651–660. doi: 10.1083/jcb.130.3.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Minet M., Nurse P., Thuriaux P., Mitchison J. M. Uncontrolled septation in a cell division cycle mutant of the fission yeast Schizosaccharomyces pombe. J Bacteriol. 1979 Jan;137(1):440–446. doi: 10.1128/jb.137.1.440-446.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moreno S., Klar A., Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. doi: 10.1016/0076-6879(91)94059-l. [DOI] [PubMed] [Google Scholar]
- Motegi F., Nakano K., Kitayama C., Yamamoto M., Mabuchi I. Identification of Myo3, a second type-II myosin heavy chain in the fission yeast Schizosaccharomyces pombe. FEBS Lett. 1997 Dec 29;420(2-3):161–166. doi: 10.1016/s0014-5793(97)01510-x. [DOI] [PubMed] [Google Scholar]
- Nagai K., Oubridge C., Ito N., Avis J., Evans P. The RNP domain: a sequence-specific RNA-binding domain involved in processing and transport of RNA. Trends Biochem Sci. 1995 Jun;20(6):235–240. doi: 10.1016/s0968-0004(00)89024-6. [DOI] [PubMed] [Google Scholar]
- Naqvi N. I., Eng K., Gould K. L., Balasubramanian M. K. Evidence for F-actin-dependent and -independent mechanisms involved in assembly and stability of the medial actomyosin ring in fission yeast. EMBO J. 1999 Feb 15;18(4):854–862. doi: 10.1093/emboj/18.4.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nash R. S., Volpe T., Futcher B. Isolation and characterization of WHI3, a size-control gene of Saccharomyces cerevisiae. Genetics. 2001 Apr;157(4):1469–1480. doi: 10.1093/genetics/157.4.1469. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nurse P., Thuriaux P., Nasmyth K. Genetic control of the cell division cycle in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1976 Jul 23;146(2):167–178. doi: 10.1007/BF00268085. [DOI] [PubMed] [Google Scholar]
- Ram A. F., Wolters A., Ten Hoopen R., Klis F. M. A new approach for isolating cell wall mutants in Saccharomyces cerevisiae by screening for hypersensitivity to calcofluor white. Yeast. 1994 Aug;10(8):1019–1030. doi: 10.1002/yea.320100804. [DOI] [PubMed] [Google Scholar]
- Roncero C., Valdivieso M. H., Ribas J. C., Durán A. Isolation and characterization of Saccharomyces cerevisiae mutants resistant to Calcofluor white. J Bacteriol. 1988 Apr;170(4):1950–1954. doi: 10.1128/jb.170.4.1950-1954.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
- Saleki R., Jia Z., Karagiannis J., Young P. G. Tolerance of low pH in Schizosaccharomyces pombe requires a functioning pub1 ubiquitin ligase. Mol Gen Genet. 1997 May 20;254(5):520–528. doi: 10.1007/s004380050447. [DOI] [PubMed] [Google Scholar]
- Salimova E., Sohrmann M., Fournier N., Simanis V. The S. pombe orthologue of the S. cerevisiae mob1 gene is essential and functions in signalling the onset of septum formation. J Cell Sci. 2000 May;113(Pt 10):1695–1704. doi: 10.1242/jcs.113.10.1695. [DOI] [PubMed] [Google Scholar]
- Sawin K. E. Cytokinesis: Sid signals septation. 2000 Jul 27-Aug 10Curr Biol. 10(15):R547–R550. doi: 10.1016/s0960-9822(00)00613-8. [DOI] [PubMed] [Google Scholar]
- Schmidt S., Sohrmann M., Hofmann K., Woollard A., Simanis V. The Spg1p GTPase is an essential, dosage-dependent inducer of septum formation in Schizosaccharomyces pombe. Genes Dev. 1997 Jun 15;11(12):1519–1534. doi: 10.1101/gad.11.12.1519. [DOI] [PubMed] [Google Scholar]
- Shu Y., Yang H., Hallberg E., Hallberg R. Molecular genetic analysis of Rts1p, a B' regulatory subunit of Saccharomyces cerevisiae protein phosphatase 2A. Mol Cell Biol. 1997 Jun;17(6):3242–3253. doi: 10.1128/mcb.17.6.3242. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sipiczki M., Bozsik A. The use of morphomutants to investigate septum formation and cell separation in Schizosaccharomyces pombe. Arch Microbiol. 2000 Dec;174(6):386–392. doi: 10.1007/s002030000214. [DOI] [PubMed] [Google Scholar]
- Sipiczki M., Grallert B., Miklos I. Mycelial and syncytial growth in Schizosaccharomyces pombe induced by novel septation mutations. J Cell Sci. 1993 Feb;104(Pt 2):485–493. doi: 10.1242/jcs.104.2.485. [DOI] [PubMed] [Google Scholar]
- Sohrmann M., Fankhauser C., Brodbeck C., Simanis V. The dmf1/mid1 gene is essential for correct positioning of the division septum in fission yeast. Genes Dev. 1996 Nov 1;10(21):2707–2719. doi: 10.1101/gad.10.21.2707. [DOI] [PubMed] [Google Scholar]
- Sohrmann M., Schmidt S., Hagan I., Simanis V. Asymmetric segregation on spindle poles of the Schizosaccharomyces pombe septum-inducing protein kinase Cdc7p. Genes Dev. 1998 Jan 1;12(1):84–94. doi: 10.1101/gad.12.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sparks C. A., Morphew M., McCollum D. Sid2p, a spindle pole body kinase that regulates the onset of cytokinesis. J Cell Biol. 1999 Aug 23;146(4):777–790. doi: 10.1083/jcb.146.4.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanaka K., Petersen J., MacIver F., Mulvihill D. P., Glover D. M., Hagan I. M. The role of Plo1 kinase in mitotic commitment and septation in Schizosaccharomyces pombe. EMBO J. 2001 Mar 15;20(6):1259–1270. doi: 10.1093/emboj/20.6.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taricani Lorena, Tejada Max L., Young Paul G. The fission yeast ES2 homologue, Bis1, interacts with the Ish1 stress-responsive nuclear envelope protein. J Biol Chem. 2001 Dec 20;277(12):10562–10572. doi: 10.1074/jbc.M110686200. [DOI] [PubMed] [Google Scholar]
- Varani G., Nagai K. RNA recognition by RNP proteins during RNA processing. Annu Rev Biophys Biomol Struct. 1998;27:407–445. doi: 10.1146/annurev.biophys.27.1.407. [DOI] [PubMed] [Google Scholar]
- Wong K. C., Naqvi N. I., Iino Y., Yamamoto M., Balasubramanian M. K. Fission yeast Rng3p: an UCS-domain protein that mediates myosin II assembly during cytokinesis. J Cell Sci. 2000 Jul;113(Pt 13):2421–2432. doi: 10.1242/jcs.113.13.2421. [DOI] [PubMed] [Google Scholar]
- Zilahi E., Miklós I., Sipiczki M. The Schizosaccharomyces pombe sep15+ gene encodes a protein homologous to the Med8 subunit of the Saccharomyces cerevisiae transcriptional mediator complex. Curr Genet. 2000 Dec;38(5):227–232. doi: 10.1007/s002940000158. [DOI] [PubMed] [Google Scholar]
- Zilahi E., Salimova E., Simanis V., Sipiczki M. The S. pombe sep1 gene encodes a nuclear protein that is required for periodic expression of the cdc15 gene. FEBS Lett. 2000 Sep 15;481(2):105–108. doi: 10.1016/s0014-5793(00)01990-6. [DOI] [PubMed] [Google Scholar]