title: Brittle Culm 12, a dual-targeting kinesin-4 protein, controls cell-cycle progression and wall properties in rice.
abstract: Kinesins are encoded by a large gene family involved in many basic processes of plant development. However, the number of functionally identified kinesins in rice is very limited. Here, we report the functional characterization of Brittle Culm12 (BC12), a gene encoding a kinesin-4 protein. bc12 mutants display dwarfism resulting from a significant reduction in cell number and brittleness due to an alteration in cellulose microfibril orientation and wall composition. BC12 is expressed mainly in tissues undergoing cell division and secondary wall thickening. In vitro biochemical analyses verified BC12 as an authentic motor protein. This protein was present in both the nucleus and cytoplasm and associated with microtubule arrays during cell division. Mitotic microtubule array comparison, flow cytometric analysis and expression assays of cyclin-dependent kinase (CDK) complexes in root-tip cells showed that cell-cycle progression is affected in bc12 mutants. BC12 is very probably regulated by CDKA;3 based on yeast two-hybrid and microarray data. Therefore, BC12 functions as a dual-targeting kinesin protein and is implicated in cell-cycle progression, cellulose microfibril deposition and wall composition in the monocot plant rice.
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Mu
authors.initials: M
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Baocai
authors.initials: B
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310006, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Qian
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Junwen
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Xiangling
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Rice Biology, China National Rice Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310006, China.
authors.forename: Dali
authors.initials: D
authors.lastname: Zeng
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jiayang
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yihua
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D002453:Cell Cycle; D002473:Cell Wall / Q000378:metabolism*; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D012275:Oryza / Q000235:genetics / Q000254:growth & development*; D010766:Phosphorylation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics
chemical_list: D010940:Plant Proteins; D018749:RNA, Plant; D016547:Kinesins
references.citation: Ambrose, J.C. and Cyr, R. (2007) The kinesin ATK5 functions in early spindle assembly in Arabidopsis. Plant Cell, 19, 226-236.
references.citation: Bannigan, A., Scheible, W.R., Lukowitz, W., Fagerstrom, C., Wadsworth, P., Somerville, C. and Baskin, T.I. (2007) A conserved role for kinesin-5 in plant mitosis. J. Cell Sci. 120, 2819-2827.
references.citation: Brady, S.T. (1985) A novel brain ATPase with properties expected for the fast axonal transport motor. Nature, 317, 73-75.
references.citation: Crowell, E.F., Bischoff, V., Desprez, T., Rolland, A., Stierhof, Y.D., Schumacher, K., Gonneau, M., Hofte, H. and Vernhettes, S. (2009) Pausing of Golgi bodies on microtubules regulates secretion of cellulose synthase complexes in Arabidopsis. Plant Cell, 21, 1141-1154.
references.citation: Dissmeyer, N., Nowack, M.K., Pusch, S., Stals, H., Inze, D., Grini, P.E. and Schnittger, A. (2007) T-loop phosphorylation of Arabidopsis CDKA;1 is required for its function and can be partially substituted by an aspartate residue. Plant Cell, 19, 972-985.
references.citation: Finn, R.D., Tate, J., Mistry, J. et al. (2008) The Pfam protein families database. Nucleic Acids Res. 36, D281-D288.
references.citation: Frey, N., Klotz, J. and Nick, P. (2009) Dynamic bridges - a calponin-domain kinesin from rice links actin filaments and microtubules in both cycling and non-cycling cells. Plant Cell Physiol. 50, 1493-1506.
references.citation: Galbraith, D.W., Harkins, K.R., Maddox, J.M., Ayres, N.M., Sharma, D.P. and Firoozabady, E. (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science, 220, 1049-1051.
references.citation: Gutierrez, R., Lindeboom, J.J., Paredez, A.R., Emons, A.M. and Ehrhardt, D.W. (2009) Arabidopsis cortical microtubules position cellulose synthase delivery to the plasma membrane and interact with cellulose synthase trafficking compartments. Nat. Cell Biol. 11, 797-806.
references.citation: Hauser, M.T., Morikami, A. and Benfey, P.N. (1995) Conditional root expansion mutants of Arabidopsis. Development, 121, 1237-1252.
references.citation: Heath, I.B. (1974) A unified hypothesis for the role of membrane bound enzyme complexes and microtubules in plant cell wall synthesis. J. Theor. Biol. 48, 445-449.
references.citation: Hoebler, C., Barry, L.D. and Delort-Laval, J. (1989) Rapid hydrolysis of plant cell wall polysaccharides by gas-liquid chromatography. J. Agric. Food. Chem. 37, 360-367.
references.citation: Horton, P., Park, K.J., Obayashi, T., Fujita, N., Harada, H., Adams-Collier, C.J. and Nakai, K. (2007) WoLF PSORT: protein localization predictor. Nucleic Acids Res. 35, 585-587.
references.citation: Inze, D. and De Veylder, L. (2006) Cell cycle regulation in plant development. Annu. Rev. Genet. 40, 77-105.
references.citation: Kirk, T.K. and Obst, J.R. (1988) Lignin determination. Methods Enzymol. 161, 87-101.
references.citation: Kong, L.J. and Hanley-Bowdoin, L. (2002) A geminivirus replication protein interacts with a protein kinase and a motor protein that display different expression patterns during plant development and infection. Plant Cell, 14, 1817-1832.
references.citation: Kwon, M., Morales-Mulia, S., Brust-Mascher, I., Rogers, G.C., Sharp, D.J. and Scholey, J.M. (2004) The chromokinesin, KLP3A, drives mitotic spindle pole separation during prometaphase and anaphase and facilitates chromatid motility. Mol. Biol. Cell, 15, 219-233.
references.citation: Lawrence, C.J., Dawe, R.K., Christie, K.R. et al. (2004) A standardized kinesin nomenclature. J. Cell Biol. 167, 19-22.
references.citation: Ledbetter, M.C. and Porter, K.R. (1963) A ‘microtubule’ in plant cell fine structure. J. Cell Biol. 19, 239-250.
references.citation: Lee, Y.M. and Kim, W. (2003) Association of human kinesin superfamily protein member 4 with BRCA2-associated factor 35. Biochem. J. 374, 497-503.
references.citation: Lee, Y.R. and Liu, B. (2004) Cytoskeletal motors in Arabidopsis. Sixty-one kinesins and seventeen myosins. Plant Physiol. 136, 3877-3883.
references.citation: Lee, Y.R., Giang, H.M. and Liu, B. (2001) A novel plant kinesin-related protein specifically associates with the phragmoplast organelles. Plant Cell, 13, 2427-2439.
references.citation: Lee, Y.R., Li, Y. and Liu, B. (2007) Two Arabidopsis phragmoplast-associated kinesins play a critical role in cytokinesis during male gametogenesis. Plant Cell, 19, 2595-2605.
references.citation: Li, Y., Qian, Q., Zhou, Y. et al. (2003) BRITTLE CULM1, which encodes a COBRA-like protein, affects the mechanical properties of rice plants. Plant Cell, 15, 2020-2031.
references.citation: Lloyd, C. (1994) Why should stationary plant cells have such dynamic microtubules? Mol. Biol. Cell, 5, 1277-1280.
references.citation: Lu, L., Lee, Y.R., Pan, R., Maloof, J.N. and Liu, B. (2005) An internal motor kinesin is associated with the Golgi apparatus and plays a role in trichome morphogenesis in Arabidopsis. Mol. Biol. Cell, 16, 811-823.
references.citation: Mazumdar, M. and Misteli, T. (2005) Chromokinesins: multitalented players in mitosis. Trends Cell Biol. 15, 349-355.
references.citation: Mazumdar, M., Sundareshan, S. and Misteli, T. (2004) Human chromokinesin KIF4A functions in chromosome condensation and segregation. J. Cell Biol. 166, 613-620.
references.citation: McDonnell, A.V., Jiang, T., Keating, A.E. and Berger, B. (2006) Paircoil2: improved prediction of coiled coils from sequence. Bioinformatics, 22, 356-358.
references.citation: Morgan, D.O. (1997) Cyclin-dependent kinases: engines, clocks, and microprocessors. Annu. Rev. Cell Dev. Biol. 13, 261-291.
references.citation: Pan, R., Lee, Y.R. and Liu, B. (2004) Localization of two homologous Arabidopsis kinesin-related proteins in the phragmoplast. Planta, 220, 156-164.
references.citation: Paredez, A.R., Somerville, C.R. and Ehrhardt, D.W. (2006) Visualization of cellulose synthase demonstrates functional association with microtubules. Science, 312, 1491-1495.
references.citation: Pien, S., Wyrzykowska, J., McQueen-Mason, S., Smart, C. and Fleming, A. (2001) Local expression of expansin induces the entire process of leaf development and modifies leaf shape. Proc. Natl Acad. Sci. USA, 98, 11812-11817.
references.citation: Reddy, A.S. and Day, I.S. (2000) The role of the cytoskeleton and a molecular motor in trichome morphogenesis. Trends Plant Sci. 5, 503-505.
references.citation: Reddy, A.S. and Day, I.S. (2001) Kinesins in the Arabidopsis genome: a comparative analysis among eukaryotes. BMC Genomics, 2, 2.
references.citation: Reddy, V.S. and Reddy, A.S. (2002) The calmodulin-binding domain from a plant kinesin functions as a modular domain in conferring Ca2+-calmodulin regulation to animal plus- and minus-end kinesins. J. Biol. Chem. 277, 48058-48065.
references.citation: Reichheld, J., Vernoux, T., Lardon, F., Van Montagu, M. and Inze, D. (1999) Specific checkpoints regulate plant cell cycle progression in response to oxidative stress. Plant J. 17, 647-656.
references.citation: Roudier, F., Fernandez, A.G., Fujita, M. et al. (2005) COBRA, an Arabidopsis extracellular glycosyl-phosphatidyl inositol-anchored protein, specifically controls highly anisotropic expansion through its involvement in cellulose microfibril orientation. Plant Cell, 17, 1749-1763.
references.citation: Sazuka, T., Aichi, I., Kawai, T., Matsuo, N., Kitano, H. and Matsuoka, M. (2005) The rice mutant dwarf bamboo shoot 1: a leaky mutant of the NACK-type kinesin-like gene can initiate organ primordia but not organ development. Plant Cell Physiol. 46, 1934-1943.
references.citation: Schindelman, G., Morikami, A., Jung, J., Baskin, T.I., Carpita, N.C., Derbyshire, P., McCann, M.C. and Benfey, P.N. (2001) COBRA encodes a putative GPI-anchored protein, which is polarly localized and necessary for oriented cell expansion in Arabidopsis. Genes Dev. 15, 1115-1127.
references.citation: Seagull, R. (1992) A quantitative electron microscopic study of changes in microtubule arrays and wall microfibril orientation during in vitro cotton fiber development. J. Cell Sci. 101, 561-577.
references.citation: Sekine, Y., Okada, Y., Noda, Y., Kondo, S., Aizawa, H., Takemura, R. and Hirokawa, N. (1994) A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally. J. Cell Biol. 127, 187-201.
references.citation: Smith, L.G. (2003) Cytoskeletal control of plant cell shape: getting the fine points. Curr. Opin. Plant Biol. 6, 63-73.
references.citation: Smith, L.G. and Oppenheimer, D.G. (2005) Spatial control of cell expansion by the plant cytoskeleton. Annu. Rev. Cell Dev. Biol. 21, 271-295.
references.citation: Somerville, C., Bauer, S., Brininstool, G. et al. (2004) Toward a systems approach to understanding plant cell walls. Science, 306, 2206-2211.
references.citation: Tanaka, K., Murata, K., Yamazaki, M., Onosato, K., Miyao, A. and Hirochika, H. (2003) Three distinct rice cellulose synthase catalytic subunit genes required for cellulose synthesis in the secondary wall. Plant Physiol. 133, 73-83.
references.citation: Umeki, N., Mitsui, T., Koike, Y. and Maruta, S. (2006a) Intermolecular cross-linking of a novel rice kinesin k16 motor domain with a photoreactive ATP derivative. J. Biochem. 139, 831-836.
references.citation: Umeki, N., Mitsui, T., Umezu, N., Kondo, K. and Maruta, S. (2006b) Preparation and characterization of a novel rice plant-specific kinesin. J. Biochem. 139, 645-654.
references.citation: Vale, R.D. (2003) The molecular motor toolbox for intracellular transport. Cell, 112, 467-480.
references.citation: Vale, R.D., Reese, T.S. and Sheetz, M.P. (1985) Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell, 42, 39-50.
references.citation: Vanstraelen, M., Torres Acosta, J.A., De Veylder, L., Inze, D. and Geelen, D. (2004) A plant-specific subclass of C-terminal kinesins contains a conserved a-type cyclin-dependent kinase site implicated in folding and dimerization. Plant Physiol. 135, 1417-1429.
references.citation: Vanstraelen, M., Inze, D. and Geelen, D. (2006a) Mitosis-specific kinesins in Arabidopsis. Trends Plant Sci. 11, 167-175.
references.citation: Vanstraelen, M., Van Damme, D., De Rycke, R., Mylle, E., Inze, D. and Geelen, D. (2006b) Cell cycle-dependent targeting of a kinesin at the plasma membrane demarcates the division site in plant cells. Curr. Biol. 16, 308-314.
references.citation: Walczak, C.E. (2003) The Kin I kinesins are microtubule end-stimulated ATPases. Mol. Cell, 11, 286-288.
references.citation: Wiedemeier, A.M., Judy-March, J.E., Hocart, C.H., Wasteneys, G.O., Williamson, R.E. and Baskin, T.I. (2002) Mutant alleles of Arabidopsis RADIALLY SWOLLEN 4 and 7 reduce growth anisotropy without altering the transverse orientation of cortical microtubules or cellulose microfibrils. Development, 129, 4821-4830.
references.citation: Wymer, C.L., Fisher, D.D., Moore, R.C. and Cyr, R.J. (1996) Elucidating the mechanism of cortical microtubule reorientation in plant cells. Cell Motil. Cytoskeleton, 35, 162-173.
references.citation: Zhong, R., Burk, D.H., Morrison, W.H. 3rd and Ye, Z.H. (2002) A kinesin-like protein is essential for oriented deposition of cellulose microfibrils and cell wall strength. Plant Cell, 14, 3101-3117.