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Genetics. Oct 2003; 165(2): 477–488.
PMCID: PMC1462777

Schizosaccharomyces pombe Ras1 effector, Scd1, interacts with Klp5 and Klp6 kinesins to mediate cytokinesis.

Abstract

Fission yeast Scd1 is an exchange factor for Cdc42 and an effector of Ras1. In a screen for scd1 interacting genes, we isolated klp5 and klp6, which encode presumptive kinesins. Klp5 and Klp6 form a complex to control the same processes, which so far include microtubule dynamics and chromosome segregation. We showed that klp5 or klp6 inactivation in combination with the scd1 deletion (scd1delta) created a synthetic temperature-dependent growth defect. Further genetic analysis demonstrated that Klp5 and Klp6 interacted specifically with the Ras1-Scd1 pathway, but not with the Ras1-Byr2 pathway. In addition, Klp5 and Klp6 can stably associate with Scd1 and Cdc42. A deletion in the Scd1 C terminus, which contains the PB1 domain, prevented Scd1 binding to Klp5/6 and caused a growth defect in Klp5/6 mutant cells that is indistinguishable from that induced by scd1delta. Analysis of the double-mutant phenotype indicated that at the nonpermissive temperature, cells failed to undergo cytokinesis efficiently. These cells contained abnormal contractile rings in which F-actin and Mid1, a key regulator of F-actin ring formation and positioning, are mispositioned and fragmented. These data suggest that Klp5/6 cooperate with the Ras1-Scd1 pathway to influence proper formation of the contractile ring for cytokinesis.

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Selected References

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  • Bähler J, Steever AB, Wheatley S, Wang Y l, Pringle JR, Gould KL, 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. [PMC free article] [PubMed]
  • Bähler J, Wu JQ, Longtine MS, Shah NG, McKenzie A, 3rd, Steever AB, Wach A, Philippsen P, Pringle JR. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast. 1998 Jul;14(10):943–951. [PubMed]
  • Barbet N, Muriel WJ, Carr AM. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene. 1992 May 1;114(1):59–66. [PubMed]
  • Bauman P, Cheng QC, Albright CF. The Byr2 kinase translocates to the plasma membrane in a Ras1-dependent manner. Biochem Biophys Res Commun. 1998 Mar 17;244(2):468–474. [PubMed]
  • Beinhauer JD, Hagan IM, Hegemann JH, Fleig U. Mal3, the fission yeast homologue of the human APC-interacting protein EB-1 is required for microtubule integrity and the maintenance of cell form. J Cell Biol. 1997 Nov 3;139(3):717–728. [PMC free article] [PubMed]
  • Chang EC, Barr M, Wang Y, Jung V, Xu HP, Wigler MH. Cooperative interaction of S. pombe proteins required for mating and morphogenesis. Cell. 1994 Oct 7;79(1):131–141. [PubMed]
  • Chang E, Bartholomeusz G, Pimental R, Chen J, Lai H, Wang L h, Yang P, Marcus S. Direct binding and In vivo regulation of the fission yeast p21-activated kinase shk1 by the SH3 domain protein scd2. Mol Cell Biol. 1999 Dec;19(12):8066–8074. [PMC free article] [PubMed]
  • Chang F. Movement of a cytokinesis factor cdc12p to the site of cell division. Curr Biol. 9(15):849–852. [PubMed]
  • 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. [PubMed]
  • Chen CR, Li YC, Chen J, Hou MC, Papadaki P, Chang EC. Moe1, a conserved protein in Schizosaccharomyces pombe, interacts with a Ras effector, Scd1, to affect proper spindle formation. Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):517–522. [PMC free article] [PubMed]
  • Forsburg SL, Sherman DA. General purpose tagging vectors for fission yeast. Gene. 1997 Jun 3;191(2):191–195. [PubMed]
  • Fukui Y, Kaziro Y. Molecular cloning and sequence analysis of a ras gene from Schizosaccharomyces pombe. EMBO J. 1985 Mar;4(3):687–691. [PMC free article] [PubMed]
  • Fukui Y, Yamamoto M. Isolation and characterization of Schizosaccharomyces pombe mutants phenotypically similar to ras1-. Mol Gen Genet. 1988 Dec;215(1):26–31. [PubMed]
  • Garcia Miguel Angel, Koonrugsa Nirada, Toda Takashi. Spindle-kinetochore attachment requires the combined action of Kin I-like Klp5/6 and Alp14/Dis1-MAPs in fission yeast. EMBO J. 2002 Nov 15;21(22):6015–6024. [PMC free article] [PubMed]
  • Garcia Miguel Angel, Koonrugsa Nirada, Toda Takashi. Two kinesin-like Kin I family proteins in fission yeast regulate the establishment of metaphase and the onset of anaphase A. Curr Biol. 2002 Apr 16;12(8):610–621. [PubMed]
  • Gilbreth M, Yang P, Wang D, Frost J, Polverino A, Cobb MH, Marcus S. The highly conserved skb1 gene encodes a protein that interacts with Shk1, a fission yeast Ste20/PAK homolog. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13802–13807. [PMC free article] [PubMed]
  • Kim HW, Yang P, Qyang Y, Lai H, Du H, Henkel JS, Kumar K, Bao S, Liu M, Marcus S. Genetic and molecular characterization of Skb15, a highly conserved inhibitor of the fission yeast PAK, Shk1. Mol Cell. 2001 May;7(5):1095–1101. [PubMed]
  • Li YC, Chen CR, Chang EC. Fission yeast Ras1 effector Scd1 interacts with the spindle and affects its proper formation. Genetics. 2000 Nov;156(3):995–1004. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Marcus S, Polverino A, Chang E, Robbins D, Cobb MH, Wigler MH. Shk1, a homolog of the Saccharomyces cerevisiae Ste20 and mammalian p65PAK protein kinases, is a component of a Ras/Cdc42 signaling module in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):6180–6184. [PMC free article] [PubMed]
  • Sutherland BW, Spiegelman GB, Weeks G. A Ras subfamily GTPase shows cell cycle-dependent nuclear localization. EMBO Rep. 2001 Nov;2(11):1024–1028. [PMC free article] [PubMed]
  • Mata J, Nurse P. tea1 and the microtubular cytoskeleton are important for generating global spatial order within the fission yeast cell. Cell. 1997 Jun 13;89(6):939–949. [PubMed]
  • Tran PT, Marsh L, Doye V, Inoué S, Chang F. A mechanism for nuclear positioning in fission yeast based on microtubule pushing. J Cell Biol. 2001 Apr 16;153(2):397–411. [PMC free article] [PubMed]
  • McCollum D, Gould KL. Timing is everything: regulation of mitotic exit and cytokinesis by the MEN and SIN. Trends Cell Biol. 2001 Feb;11(2):89–95. [PubMed]
  • Van Aelst L, D'Souza-Schorey C. Rho GTPases and signaling networks. Genes Dev. 1997 Sep 15;11(18):2295–2322. [PubMed]
  • Murray JM, Johnson DI. The Cdc42p GTPase and its regulators Nrf1p and Scd1p are involved in endocytic trafficking in the fission yeast Schizosaccharomyces pombe. J Biol Chem. 2001 Feb 2;276(5):3004–3009. [PubMed]
  • Verde F, Wiley DJ, Nurse P. Fission yeast orb6, a ser/thr protein kinase related to mammalian rho kinase and myotonic dystrophy kinase, is required for maintenance of cell polarity and coordinates cell morphogenesis with the cell cycle. Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7526–7531. [PMC free article] [PubMed]
  • Murray JM, Johnson DI. Isolation and characterization of Nrf1p, a novel negative regulator of the Cdc42p GTPase in Schizosaccharomyces pombe. Genetics. 2000 Jan;154(1):155–165. [PMC free article] [PubMed]
  • Vojtek AB, Hollenberg SM, Cooper JA. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. [PubMed]
  • Wang Y, Xu HP, Riggs M, Rodgers L, Wigler M. byr2, a Schizosaccharomyces pombe gene encoding a protein kinase capable of partial suppression of the ras1 mutant phenotype. Mol Cell Biol. 1991 Jul;11(7):3554–3563. [PMC free article] [PubMed]
  • Papadaki Piyi, Pizon Véronique, Onken Brian, Chang Eric C. Two ras pathways in fission yeast are differentially regulated by two ras guanine nucleotide exchange factors. Mol Cell Biol. 2002 Jul;22(13):4598–4606. [PMC free article] [PubMed]
  • West RR, Malmstrom T, Troxell CL, McIntosh JR. Two related kinesins, klp5+ and klp6+, foster microtubule disassembly and are required for meiosis in fission yeast. Mol Biol Cell. 2001 Dec;12(12):3919–3932. [PMC free article] [PubMed]
  • Prokopenko SN, Saint R, Bellen HJ. Untying the Gordian knot of cytokinesis. Role of small G proteins and their regulators. J Cell Biol. 2000 Mar 6;148(5):843–848. [PMC free article] [PubMed]
  • West Robert R, Malmstrom Terra, McIntosh J Richard. Kinesins klp5(+) and klp6(+) are required for normal chromosome movement in mitosis. J Cell Sci. 2002 Mar 1;115(Pt 5):931–940. [PubMed]
  • Qyang Yibing, Yang Peirong, Du Hongyan, Lai Hong, Kim HyeWon, Marcus Stevan. The p21-activated kinase, Shk1, is required for proper regulation of microtubule dynamics in the fission yeast, Schizosaccharomyces pombe. Mol Microbiol. 2002 Apr;44(2):325–334. [PubMed]
  • Yen Hsueh-Chi S, Gordon Colin, Chang Eric C. Schizosaccharomyces pombe Int6 and Ras homologs regulate cell division and mitotic fidelity via the proteasome. Cell. 2003 Jan 24;112(2):207–217. [PubMed]
  • Sawin KE, Nurse P. Regulation of cell polarity by microtubules in fission yeast. J Cell Biol. 1998 Jul 27;142(2):457–471. [PMC free article] [PubMed]

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