Abstract
As a cell cycle regulator, the Myb-related CDC5 protein was reported to be essential for the G2 phase of the cell cycle in yeast and animals, but little is known about its function in plants. Here we report the functional characterization of the CDC5 gene in Arabidopsis thaliana. Arabidopsis CDC5 (AtCDC5) is mainly expressed in tissues with high cell division activity, and is expressed throughout the entire process of embryo formation. The AtCDC5 loss-of-function mutant is embryonic lethal. In order to investigate the function of AtCDC5 in vivo, we generated AtCDC5-RNAi plants in which the expression of AtCDC5 was reduced by RNA interference. We found that the G2 to M (G2/M) phase transition was affected in the AtCDC5-RNAi plants, and that endoreduplication was increased. Additionally, the maintenance of shoot apical meristem (SAM) function was disturbed in the AtCDC5-RNAi plants, in which both the WUSCHEL (WUS)-CLAVATA (CLV) and the SHOOT MERISTEMLESS (STM) pathways were impaired. In situ hybridization analysis showed that the expression of STM was greatly reduced in the shoot apical cells of the AtCDC5-RNAi plants. Moreover, cyclinB1 or Histone4 was found to be expressed in some of these cells when the transcript of STM was undetectable. These results suggest that AtCDC5 is essential for the G2/M phase transition and may regulate the function of SAM by controlling the expression of STM and WUS.
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References
Brodsky VY, Uryvaeva IV . Cell polyploidy: its relation to tissue growth and function. Int Rev Cytol 1977; 50:275–332.
D'Amato F . Endopolyploidy as a factor in plant tissue development. Caryologia 1964; 17:41–52.
Boudolf V, Barrôco R, Engler Jde A, et al. B1-type cyclin-dependent kinases are essential for the formation of stomatal complexes in Arabidopsis thaliana. Plant Cell 2004; 16:945–955.
Boudolf V, Vlieghe K, Beemster GT, et al. The plant-specific cyclin-dependent kinase CDKB1;1 and transcription factor E2Fa-DPa control the balance of mitotically dividing and endoreduplicating cells in Arabidopsis. Plant Cell 2004; 16:2683–2692.
Weinl C, Marquardt S, Kuijt SJ, et al. Novel functions of plant cyclin-dependent kinase inhibitors, ICK1/KRP1, can act non-cell-autonomously and inhibit entry into mitosis. Plant Cell 2005; 17:1704–1722.
Schnittger A, Weinl C, Bouyer D, Schobinger U, Hülskamp M . Misexpression of the cyclin-dependent kinase inhibitor ICK1/KRP1 in single-celled Arabidopsis trichomes reduces endoreduplication and cell size and induces cell death. Plant Cell 2003; 15:303–315.
Sugimoto-Shirasu K, Roberts K . “Big it up”: endoreduplication and cell-size control in plants. Curr Opin Plant Biol 2003; 6:544–553.
Barton MK, Poethig RS . Formation of the shoot apical meristem in Arabidopsis thaliana: an analysis of development in the wild type and in the shoot meristemless mutant. Development 1993; 119:823–831.
Clark SE, Jacobsen SE, Levin JZ, Meyerowitz EM . The CLAVATA and SHOOT MERISTEMLESS loci competitively regulate meristem activity in Arabidopsis. Development 1996; 122:1567–1575.
Endrizzi K, Moussian B, Haecker A, Levin JZ, Laux T . The SHOOT MERISTEMLESS gene is required for maintenance of undifferentiated cells in Arabidopsis shoot and floral meristems and acts at a different regulatory level than the meristem genes WUSCHEL and ZWILLE. Plant J 1996; 10:967–979.
Laux T, Mayer KFX, Berger J, Jürgens G . The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development 1996; 122:87–96.
Long J, Moan E, Medford J, Barton M . A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature 1996; 379:66–69.
Schoof H, Lenhard M, Haecker A, Mayer KFX, Jurgens G, Laux T . The stem cell population of Arabidopsis shoot meristems is maintained by a regulatory loop between the CLAVATA and WUSCHEL genes. Cell 2000; 100:635–644.
Trotochaud A, Jeong S, Clark S . CLAVATA3, a multimeric ligand for the CLAVATA1 receptor-kinase. Science 2000; 289:613–617.
Byrne M, Barley R, Curtis M, et al. Asymmetric leaves1 mediates leaf patterning and stem cell function in Arabidopsis. Nature 2000; 408:967–971.
Iwakawa H, Ueno Y, Semiarti E, et al. The ASYMMETRIC LEAVES2 gene of Arabidopsis thaliana, required for formation of a symmetric flat leaf lamina, encodes a member of a novel family of proteins characterized by cysteine repeats and a leucine zipper. Plant Cell Physiol 2002; 43:467–478.
Ori N, Eshed Y, Chuck G, Bowman JL, Hake S . Mechanisms that control knox gene expression in the Arabidopsis shoot. Development 2000; 127:5523–5532.
Semiarti E, Ueno Y, Tsukaya H, Iwakawa H, Machida C, Machida Y . The asymmetric leaves2 gene of Arabidopsis thaliana regulates formation of a symmetric lamina, establishment of venation and repression of meristem-related homeobox genes in leaves. Development 2001; 128:1771–1783.
de Jager SM, Maughan S, Dewitte W, Scofield S, Murray JA . The developmental context of cell-cycle control in plants. Semin Cell Dev Biol 2005; 16:385–396.
Ohi R, Feoktistova A, McCann S, et al. Myb-related Schizosaccharomyces pombe cdc5p is structurally and functionally conserved in eukaryotes. Mol Cell Biol 1998; 18:4097–4108.
Ohi R, McCollum D, Hirani B, et al. The Schizosaccharomyces pombe cdc5+ gene encodes an essential protein with homology to c-Myb. EMBO J 1994; 13:471–483.
Bernstein HS, Coughlin SR . Pombe Cdc5-related protein. A putative human transcription factor implicated in mitogen-activated signaling. J Biol Chem 1997; 272:5833–5837.
Bernstein HS, Coughlin SR . A mammalian homolog of fission yeast Cdc5 regulates G2 progression and mitotic entry. J Biol Chem 1998; 273:4666–4671.
Hirayama T, Shinozaki K . A cdc5+ homolog of a higher plant, Arabidopsis thaliana. Proc Natl Acad Sci USA 1996; 93:13371–13376.
Lei X, Shen X, Xu X, Bernstein H . Human Cdc5, a regulator of mitotic entry, can act as a site-specific DNA binding protein. J Cell Sci 2000; 113:4523–4531.
Ajuh P, Kuster B, Panov K, Zomerdijk JC, Mann M, Lamond AI . Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry. EMBO J 2000; 19:6569–6581.
Burns CG, Ohi R, Krainer AR, Gould KL . Evidence that Myb-related CDC5 proteins are required for pre-mRNA splicing. Proc Natl Acad Sci USA 1999; 96:13789–13794.
Liu L, Graub R, Hlaing M, et al. Distinct domains of human CDC5 direct its nuclear import and association with the spliceosome. Cell Biochem Biophys 2003; 39:119–132.
McDonald WH, Ohi R, Smelkova N, Frendewey D, Gould KL . Myb-related fission yeast cdc5p is a component of a 40S snRNP-containing complex and is essential for pre-mRNA splicing. Mol Cell Biol 1999; 19:5352–5362.
Tsai WY, Chow YT, Chen HR, et al. Cef1p is a component of the Prp19p-associated complex and essential for pre-mRNA splicing. J Biol Chem 1999; 274:9455–9462.
Burns CG, Ohi R, Mehta S, et al. Removal of a single alpha-tubulin gene intron suppresses cell cycle arrest phenotypes of splicing factor mutations in Saccharomyces cerevisiae. Mol Cell Biol 2002; 22:801–815.
Dahan O, Kupiec M . Mutations in genes of Saccharomyces cerevisiae encoding pre-mRNA splicing factors cause cell cycle arrest through activation of the spindle checkpoint. Nucleic Acids Res 2002; 30:4361–4370.
Lin Z, Yin K, Wang X, et al. Virus induced gene silencing of AtCDC5 results in accelerated cell death in Arabidopsis leaves. Plant Physiol Biochem 2007; 45:87–94.
Chuang CF, Meyerowitz EM . Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci USA 2000; 97:4985–4990.
Clough SJ, Bent AF . Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 1998; 16:735–743.
Qin GJ, Gu HY, Zhao YD, et al. An indole-3-acetic acid carboxyl methyltransferase regulates Arabidopsis leaf development. Plant Cell 2005; 17:2693–2704.
Qu LJ, Chen J, Liu MH, et al. Molecular cloning and functional analysis of a novel type of Bowman-Birk inhibitor gene family in rice. Plant Physiol 2003; 133:560–570.
Guo L, Wang ZY, Lin H, et al. Expression and functional analysis of the rice plasma-membrane intrinsic protein gene family. Cell Res 2006; 16:277–286.
Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E . Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 1983; 220:1049–1051.
Liu CM, Meinke DW . The titan mutants of Arabidopsis are disrupted in mitosis and cell cycle control during seed development. Plant J 1998; 16:21–31.
Brand U, Grunewald M, Hobe M, Simon R . Regulation of CLV3 expression by two homeobox genes in Arabidopsis. Plant Physiol 2002; 129:565–575.
Porceddu A, Stals H, Reichheldt J, et al. A plant-specific cyclin-dependent kinase is involved in the control of G(2)/M progression in plants. J Biol Chem 2001; 276:36354–36360.
Ferreira PC, Hemerly AS, Engler JD, van Montagu M, Engler G, Inze D . Developmental expression of the Arabidopsis cyclin gene cyc1At. Plant Cell 1994; 6:1763–1774.
Hemerly A, Bergounioux C, Vanmontagu M, Inze D, Ferreira P . Genes regulating the plant cell cycle isolation of a mitotic-like cyclin from Arabidopsis thaliana. Proc Natl Acad Sci USA 1992; 89:3295–3299.
Wang CX, Liu ZC . Arabidopsis ribonucleotide reductases are critical for cell cycle progression, DNA damage repair, and plant development. Plant Cell 2006; 18:350–365.
Zimmermann P, Hirsch-Hoffmann M, Hennig L, Gruissem W . GENEVESTIGATOR. Arabidopsis microarray database and analysis toolbox. Plant Physiol 2004; 136:2621–2632.
Potter CJ, Xu T . Mechanisms of size control. Curr Opin Genet Dev 2001; 11:279–286.
Weigmann K, Cohen SM, Lehner CF . Cell cycle progression, growth and patterning in imaginal discs despite inhibition of cell division after inactivation of Drosophila Cdc2 kinase. Development 1997; 124:3555–3563.
Tsukaya H . Organ shape and size: a lesson from studies of leaf morphogenesis. Curr Opin Plant Biol 2003; 6:57–62.
Gross-Hardt R, Laux T . Stem cell regulation in the shoot meristem. J Cell Sci 2003; 116:1659–1666.
Acknowledgements
The authors thank Dr Liying Du (Peking University, China) for technical help on the flow cytometric analysis. The authors also thank Dr Zhongchi Liu (University of Maryland, USA), Dr Chun-Ming Liu (Institute of Botany CAS, China), Dr Terry Matthew (University of Southampton, UK), Professor Daochun Kong (Peking University, China) and Dr Naomi Nakayama (Yale University, USA) for critical comments and valuable discussion. This work was supported by the National Natural Science Foundation of China (GN 30625002 to L-J Qu).
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Lin, Z., Yin, K., Zhu, D. et al. AtCDC5 regulates the G2 to M transition of the cell cycle and is critical for the function of Arabidopsis shoot apical meristem. Cell Res 17, 815–828 (2007). https://doi.org/10.1038/cr.2007.71
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DOI: https://doi.org/10.1038/cr.2007.71
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