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Characterization of in vivo functions of Nicotiana benthamiana RabE1

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Abstract

We characterized the gene expression, subcellular localization, and in vivo functions of a Nicotiana benthamiana small GTPase belonging to the RabE family, designated NbRabE1. The NbRabE1 promoter drove strong β-glucuronidase reporter expression in young tissues containing actively dividing cells and in stomata guard cells. GFP fusion proteins of NbRabE1 and its dominant-negative and constitutively active mutants were all localized to the Golgi apparatus and the plasma membrane but showed different affinities for membrane attachment. Virus-induced gene silencing of NbRabE1 resulted in pleiotropic phenotypes, including growth arrest, premature senescence, and abnormal leaf development. At the cellular level, the leaves in which NbRabE1 was silenced contained abnormal stomata that lacked pores or contained incomplete ventral walls, suggesting that NbRabE1 deficiency leads to defective guard cell cytokinesis. Ectopic expression of the dominant-negative mutant of NbRabE1 in Arabidopsis thaliana resulted in retardation of shoot and root growth accompanied by defective root hair formation. These developmental defects are discussed in conjunction with proposed functions of RabE GTPases in polarized secretory vesicle trafficking.

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References

  • Ahn CS, Han J-A, Lee H-S, Lee S, Pai H-S (2011) The PP2A regulatory subunit Tap46, a component of TOR signaling pathway, modulates growth and metabolism in plants. Plant Cell 23:185–209

    Article  PubMed  CAS  Google Scholar 

  • Bergmann DC, Sack FD (2007) Stomatal development. Annu Rev Plant Biol 58:163–181

    Article  PubMed  CAS  Google Scholar 

  • Bogdanove AJ, Martin GB (2000) AvrPto-dependent Pto-interacting proteins and AvrPto-interacting proteins in tomato. PNAS 97:8836–8840

    Article  PubMed  CAS  Google Scholar 

  • Boguski MS, McCormick F (1993) Proteins regulating Ras and its relatives. Nature 366:643–654

    Article  PubMed  CAS  Google Scholar 

  • Bolte S, Talbot C, Boutte Y, Catrice O, Read ND, Satiat-Jeunemaitre B (2004) FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells. J Microsc 214:159–173

    Article  PubMed  CAS  Google Scholar 

  • Carter CJ, Bednarek SY, Raikhel NV (2004) Membrane trafficking in plants: new discoveries and approaches. Curr Opin Plant Biol 7:701–707

    Article  PubMed  CAS  Google Scholar 

  • Cho HS, Lee SS, Kim KD, Kim SJ, Hwang I, Lim JS, Park YI, Pai HS (2004) DNA gyrase is involved in chloroplast nucleoid partitioning. Plant Cell 16:2665–2682

    Article  PubMed  CAS  Google Scholar 

  • Falbel TG, Koch LM, Nadeau JA, Segui-Simarro JM, Sack FD, Bednarek SY (2003) SCD1 is required for cell cytokinesis and polarized cell expansion in Arabidopsis thaliana. Development 130:4011–4024

    Article  PubMed  CAS  Google Scholar 

  • Fendrych M, Synek L, Pečenková T, Toupalová H, Cole R, Drdová E, Nebesářová J, Šedinová M, Hála M, Fowler JE, Žárský V (2010) The Arabidopsis exocyst complex is involved in cytokinesis and cell plate maturation. Plant Cell 22:3053–3065

    Article  PubMed  CAS  Google Scholar 

  • Goh T, Uchida W, Arakawa S, Ito E, Dainobu T, Ebine K, Takeuchi M, Sato K, Ueda T, Nakano A (2007) VPS9a, the Common Activator for Two Distinct Types of Rab5 GTPases, Is Essential for the Development of Arabidopsis thaliana. Plant Cell 19:3504–3515

    Article  PubMed  CAS  Google Scholar 

  • Goud B, Salminen A, Walworth NC, Novick PJ (1988) A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast. Cell 53:753–768

    Article  PubMed  CAS  Google Scholar 

  • Haga N, Kato K, Murase M, Araki S, Kubo M, Demura T, Suzuki K, Müller I, Voß U, Jürgens G, Ito M (2007) R1R2R3-Myb proteins positively regulate cytokinesis through activation of KNOLLE transcription in Arabidopsis thaliana. Development 134:1101–1110

    Article  PubMed  CAS  Google Scholar 

  • Hattula K, Furuhjelm J, Tikkanen J, Tanhuanpää K, Laakkonen P, Peränen J (2006) Characterization of the Rab8-specific membrane traffic route linked to protrusion formation. J Cell Sci 119:4866–4877

    Article  PubMed  CAS  Google Scholar 

  • Huber LA, Pimplikar S, Parton RG, Virta H, Zerial M, Simons K (1993) Rab8, a small GTPase involved in vesicular traffic between the TGN and the basolateral plasma membrane. J Cell Biol 123:35–45

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    PubMed  CAS  Google Scholar 

  • Jin H, Axtell MJ, Dahlbeck D, Ekwenna O, Zhang S, Staskawicz B, Baker B (2002) NPK1, an MEKK1-like mitogen-activated protein kinase kinase kinase, regulates innate immunity and development in plants. Dev Cell 3:291–297

    Article  PubMed  CAS  Google Scholar 

  • Kim M, Lim JH, Ahn CS, Park K, Kim GT, Kim WT, Pai HS (2006) Mitochondria-associated hexokinases play a role in the control of programmed cell death in Nicotiana benthamiana. Plant Cell 18:2341–2355

    Article  PubMed  CAS  Google Scholar 

  • Lee JY, Lee HS, Wi SJ, Park KY, Schmit AC, Pai HS (2009) Dual functions of Nicotiana benthamiana Rae1 in interphase and mitosis. Plant J 59:278–291

    Article  PubMed  CAS  Google Scholar 

  • Marat AL, Dokainish H, McPherson PS (2011) DENN domain proteins: regulators of Rab GTPases. J Biol Chem 286:13791–13800

    Article  PubMed  CAS  Google Scholar 

  • Nadeau JA, Sack FD (2002) Control of stomatal distribution on the Arabidopsis leaf surface. Science 296:1697–1700

    Article  PubMed  CAS  Google Scholar 

  • Nelson BK, Cai X, Nebenfuhr A (2007) A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants. Plant J 51:1126–1136

    Article  PubMed  CAS  Google Scholar 

  • Nielsen E, Cheung AY, Ueda T (2008) The regulatory RAB and ARF GTPases for vesicular trafficking. Plant Physiol 147:1516–1526

    Article  PubMed  CAS  Google Scholar 

  • Nishihama R, Ishikawa M, Araki S, Soyano T, Asada T, Machida Y (2001) The NPK1 mitogen-activated protein kinase kinase kinase is a regulator of cell-plate formation in plant cytokinesis. Genes Dev 15:352–363

    Article  PubMed  CAS  Google Scholar 

  • Ponnambalam S, Baldwin SA (2003) Constitutive protein secretion from the trans-Golgi network to the plasma membrane. Mol Membrane Biol 20:129–139

    Article  CAS  Google Scholar 

  • Rojo E, Gillmor CS, Kovaleva V, Somerville CR, Raikhel NV (2001) VACUOLELESS1 is an essential gene required for vacuole formation and morphogenesis in Arabidopsis. Dev Cell 1:303–310

    Article  PubMed  CAS  Google Scholar 

  • Ryan E, Steer M, Dolan L (2001) Cell biology and genetics of root hair formation in Arabidopsis thaliana. Protoplasma 215:140–149

    Article  PubMed  CAS  Google Scholar 

  • Salminen A, Novick PJ (1987) A ras-like protein is required for a post-Golgi event in yeast secretion. Cell 49:527–538

    Article  PubMed  CAS  Google Scholar 

  • Sato T, Mushiake S, Kato Y, Sato K, Sato M, Takeda N, Ozono K, Miki K, Kubo Y, Tsuji A, Harada R, Harada A (2007) The Rab8 GTPase regulates apical protein localization in intestinal cells. Nature 448:366–369

    Article  PubMed  CAS  Google Scholar 

  • Schiefelbein JW (2000) Constructing a plant cell: the genetic control of root hair development. Plant Physiol 124:1525–1531

    Article  PubMed  CAS  Google Scholar 

  • Söllner R, Glässer G, Wanner G, Somerville CR, Jürgens G, Farhah F, Assaad FF (2002) Cytokinesis-defective mutants of Arabidopsis. Plant Physiol 129:678–690

    Article  PubMed  Google Scholar 

  • Speth EB, Imboden L, Hauck P, He SY (2009) Subcellular localization and functional analysis of the Arabidopsis GTPase RabE. Plant Physiol 149:1824–1837

    Article  PubMed  CAS  Google Scholar 

  • Stenmark H, Olkkonen VM (2001) The Rab GTPase family. Genome Biol 2:3007.1–3007.7

    Article  Google Scholar 

  • Takai Y, Sasaki T, Matozaki T (2001) Small GTP-binding proteins. Physiol Rev 81:153–208

    PubMed  CAS  Google Scholar 

  • Vernoud V, Horton AC, Yang Z, Nielsen E (2003) Analysis of the small GTPase gene superfamily of Arabidopsis. Plant Physiol 131:1191–1208

    Article  PubMed  CAS  Google Scholar 

  • Voinnet O, Rivas S, Mestre P, Baulcombe D (2003) An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33:949–956

    Article  PubMed  CAS  Google Scholar 

  • Woollard AAD, Moore I (2008) The functions of Rab GTPases in plant membrane traffic. Curr Opin Plant Biol 11:610–619

    Article  PubMed  CAS  Google Scholar 

  • Yang M, Nadeau JA, Zhao L, Sack FD (1999) Characterization of a cytokinesis defective (cyd1) mutant of Arabidopsis. J Exp Bot 50:1437–1446

    PubMed  CAS  Google Scholar 

  • Zerial M, McBride H (2001) Rab proteins as membrane organizers. Nat Rev Mol Cell Biol 2:107–117

    Article  PubMed  CAS  Google Scholar 

  • Zheng H, Camacho L, Wee E, Batoko H, Legen J, Leaver CJ, Malhó R, Hussey PJ, Moore I (2005) A Rab-E GTPase mutant acts downstream of the Rab-D subclass in biosynthetic membrane traffic to the plasma membrane in tobacco leaf epidermis. Plant Cell 17:2020–2036

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the Next-Generation BioGreen 21 Program (PMBC, No. PJ009079/SSAC, No. PJ008214; RDA) and the Mid-career Researcher Program (No. 20110027612; NRF/MEST) of Korea.

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Correspondence to Hyun-Sook Pai.

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C.S. Ahn and J.-A. Han contributed equally to this work.

Genbank accession number: JQ519194 (NbRabE1).

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Ahn, C.S., Han, JA. & Pai, HS. Characterization of in vivo functions of Nicotiana benthamiana RabE1. Planta 237, 161–172 (2013). https://doi.org/10.1007/s00425-012-1760-5

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