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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (10): 1756-1766.

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Characterization and Functional Analysis of a Small GTP-binding Protein AtRAB Interacting with H+-Pyrophosphatase AVP1 in Arabidopsis thaliana

LIU Rong-Bang1,2,CHEN Ming2,*,GUO Meng-Meng2,SI Qing-Lin2,GAO Shi-Qing3,XU Zhao-Shi2,LI Lian-Cheng2,MA You-Zhi2,YIN Jun1,*   

  1. 1 Henan Agricultural University / National Key Laboratory of Wheat and Maize Crop Science / Collaborative Innovation Center of Henan Grain Crops, Zhengzhou 450002, China; 2 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture, Beijing 100081, China; 3 Beijing Hybrid Wheat Engineering and Technology Research Center, Beijing 100097, China
  • Received:2014-03-10 Revised:2014-06-16 Online:2014-10-12 Published:2014-07-23
  • Contact: 尹钧, E-mail: xmzxyj@126.com, Tel: 0371-63558203; 陈明, E-mail: chenming02@caas.cn, Tel: 010-82108789

Abstract:

H+-Pyrophosphatase (H+-PPase) is an important proton transporter in plants. It cooperates with H+-ATPase and transport protons in vacuole or extracellular area to maintain a constant H+ gradient, which enables the transport of ions and other components (e.g. amino acids, carbohydrates). In the current research, AVP1 was applied to membrane proteins-based yeast two-hybrid system, and a small GTP-binding protein AtRAB was identified by screening the Arabidopsis cDNA library. The interaction between AVP1 and AtRAB was confirmed by interaction analysis in yeast. Bimolecular fluorescent complementation (BiFC) analysis suggested that AtRAB and AVP1 interaction took place in the plasma membrane and the nucleus. Phenotypes of wild type (WT) and Arabidopsis mutants of avp1 (AVP1) and rab (AtRAB) were compared under high-salt, low-phosphorus and low-potassium conditions. Under the high salt stress, avp1 and rab presented similar phenotypes along with the increase of NaCl concentration, which were characterized by shortened root length and reduced lateral root number in comparison with the WT. The decreasing phosphorus concentration also led to the phenotypes of shortened root length, reduced lateral root number and reduced total root area in the two mutants. Particularly, stronger response was observed in rab than in avp1 under the same phosphorus concentration. The potassium treatments resulted in similar phenotypes to those in phosphorus treatment. The results indicated that AVP1 can interact with AtRAB on the plasma membrane and the nucleus and further influence the absorption of ions in plants. The two mutants of avp1 and rab showed similar phenotypes, suggesting that both AVP1 and AtRAB positively regulate plant response to high salt, low phosphorus and low potassium stresses in the same signaling pathway.

Key words: H+-Pyrophosphatase, Small GTP-binding protein AtRAB, Yeast two-hybrid system, Protein interaction, stresses response

[1]Rea P A, Poole R J. Vacuolar H+-translocating pyrophosphatase. Ann Rev Plant Physiol, 1993, 44: 157–180



[2]Nakanishi Y, Matsuda N, Aizawa K, Kashiyama T, Yamamoto K, Mimura T, Ikeda M, Maeshima M. Molecular cloning and sequencing of the cDNA for vacuolar H+-pyrophosphatase from Chara coralline. Biochim Biophys Acta, 1999, 1418: 245–250



[3]Baykov A A, Bakuleva N P, Rea P A. Steady-state kinetics of substrate hydrolysis by vacuolar H+-pyrophosphatase. A simple three-state model. Eur J Biochem, 1993, 217: 755–762



[4]Long A R, Williams L E, Nelson S J, Hall J L. Localization of membrane pyrophosphatase activity in Ricinus communis seedlings. J Plant Physiol, 1995, 146: 629–638



[5]Robinson D G, Haschke H P, Hinz G, Hoh B, Maeshima M, Marty F. Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons. Planta, 1996, 198: 95–103



[6]Robinson D G, Hoppenrath M, Oberbeck K, Luykx P, Ratajczak R. Localization of pyrophosphatase and V-ATPase in Chlamydomonas reinhardtii. Bot Acta, 1998, 111: 108–122



[7]Lerchl J, Geigenberger P, Stitt M, Sonnewald U. Impaired photoassimilate partitioning caused by phloem-specific removal of pyrophosphate can be complemented by a phloem-specific cytosolic yeast-derived invertase in transgenic plants. Plant Cell, 1995, 7: 259–270



[8]Jose R, Castineira P, Hernandez A, Drake R, Serrano. A plant proton-pumping inorganic pyrophosphatase functionally complements the vacuolar ATPase transport activity and confers bafilomycin resistance in yeast. Biochem J, 2011, 437: 269–278



[9]Blumwald E. Tonopast vesicles for the study of ion transport in plant vacuoles. Plant, 1987, 69: 731–734



[10]Hedrich R, Schroeder J I. The physiology of ion channels and electrogenic pumps in higher plants. Ann Rev Plant Physiol, 1989, 40: 539–569



[11]Guo S L, Yin H B, Zhang X, Zhao F Y, Li P H, Chen S H, Zhao Y X, Zhang H. Molecular cloning and characterization of a vacuolar H+-pyrophosphatase gene, SsVP, from the halophyte Suaeda salsa and its overexpression increases salt and drought tolerance of Arabidopsis. Plant Mol Biol, 2006, 60: 41–50



[12]Sun Q H, Gao F, Zhao L, Li K P, Zhang J R. Identification of a new 130 bp cis-acting element in the TsVP1 promoter involved in the salt stress response from Thellungiella halophila. BMC Plant Biol, 2010, 10: 1471–2290



[13]Liu L, Wang Y, Wang N, Dong Y Y, Fan X D, Liu X M, Yang J, Li H Y. Cloning of a Vacuolar H+-pyrophosphatase gene from the halophyte Suaeda corniculata whose heterogonous overexpression improves salt, saline-alkali and drought tolerance in Arabidopsis. J Integr Plant Biol, 2011, 53: 731–742



[14]Bhaskaran S, Savithramma D L. Co-expression of Pennisetum glaucum vacuolar Na+/H+ antiporter and Arabidopsis H+-pyrophosphatase enhances salt tolerance in transgenic tomato. J Exp Bot, 2011, 62: 5561–5570



[15]Park S H, Li J S, Pittman J K, Berkowitz G A, Yang H B, Undurraga S, Morris J, Hirschi K D, Gaxiola R A. Up-regulation of a H+-pyrophosphatase (H+-PPase) as a strategy to engineer drought-resistant crop plants. PNAS, 2005, 102: 18830–18835



[16]Zhang H, Shen G X, Kuppu S, Gaxiola R, Payton P. Creating drought- and salt-tolerant cotton by overexpressing a vacuolar pyrophosphatase gene. Plant Signal Behav, 2011, 6:6: 861–863



[17]Pasapula V, Shen G X, Kuppu S, Paez-Valencia J, Mendoza M, Hou P, Chen J, Qiu X Y, Zhu L F, Zhang X L, Auld D, Blumwald E, Zhang H, Gaxiola R, Payton P. Expression of an Arabidopsis vacuolar H+-pyrophosphatase gene (AVP1) in cotton improves drought-and salt tolerance and increases fibre yield in the field conditions. Plant Biotechnol J, 2011, 9: 88–99



[18]Dong Q L, Liu D D, An X H, Hu D G, Yao Y X, Hao Y J. MdVHP1 encodes an apple vacuolar H+-PPase and enhances stress tolerance in transgenic apple callus and tomato. J Plant Physiol, 2011, 168: 2124–2133



[19]Zhang J, Li J Q, Wang X C, Chen J. OVP1, a Vacuolar H+-translocating inorganic pyrophosphatase (V-PPase), overexpression improved rice cold tolerance. Plant Physiology and Biochemistry, 2011, 49: 33–38



[20]Kabala K, Janicka-Russak M, Klobus G. Different responses of tonoplast proton pumps in cucumber roots to cadmium and copper. J Plant Physiol, 2010, 167: 1328–1335



[21]Migocka M, Papierniak A, Kosatka E, Klobus G. Comparative study of the active cadmium efflux systems operating at the plasma membrane and tonoplast of cucumber root cells. J Exp Bot, 2011, 62: 4903–4916



[22]Khoudi H, Maatar Y, Gouiaa S, Masmoudi K. Transgenic tobacco plants expressing ectopically wheat H+-pyrophosphatase (H+-PPase) gene TaVP1 show enhanced accumulation and tolerance to cadmium. J Plant Physiol, 2012, 169: 98–103



[23]Li J S, Yang H B, Peer W A, Richter G, Blakeslee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S, Khodakovskaya M, Richards E L, Krizek B, Murphy A S, Gilroy S, Gaxiola R. Arabidopsis H+-PPase AVP1 regulates Auxin-mediated organ development. Science, 2005, 310: 121–125



[24]Yao Y X, Dong Q L, You C X, Zhai H, Hao Y J. Expression analysis and functional characterization of apple MdVHP1 gene reveals its involvement in Na+, malate and soluble sugar accumulation. Plant Physiol Biochem, 2011, 49: 1201–1208



[25]Krebs M, Beyhl D, Gorlich E, Al-Rasheid K A S, Marten I, Stierhof Y D, Hedrich R, Schumacher K. Arabidopsis V-ATPase activity at the tonoplast is required for efficient nutrient storage but not for sodium accumulation. Proc Natl Acad Sci USA, 2010, 107: 3251–3256



[26]Yang H B, Knapp J, Koirala P, Rajagopal D, Peer W A, Silbart L K, Murphy A, Gaxiola R A. Enhanced phosphorus nutrition in monocots and dicots over-expressing a phosphorus-responsive type I H+-pyrophosphatase. Plant Biotechnol J, 2007, 5: 735–745



[27]Stagljar I, Korostensky C, Johnsson N, te Heesen S. A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo. Proc Natl Acad Sci USA, 1998, 95: 5187–5192



[28]李敏, 杨双, 阮燕晔, 樊金娟, 张立军. 拟南芥T DNA插入突变体atsuc3的PCR鉴定. 植物生理学通讯, 2006, 42: 91–94



Li M, Yang S, Ruan Y Y, Fan J J, Zhang L J. Identification of atsuc3 with T-DNA Insertion by PCR. Plant Physiol Commun, 2006, 42: 91–94 (in Chinese)



[29]Waizenegger I, Lukowitz W, Assaad F, Schwarz H, Jurgens G, Mayer U. The Arabidopsis KNOLLE and KEULE genes interact to promote vesicle fusion during cytokinesis. Curr Biol, 2000, 10: 1371–1374



[30]Geldner N, Friml J, Stierhof Y D, Jurgens G, Palme K. Auxin transport inhibitors block PIN1 cycling and vesicle trafficking. Nature, 2001, 413: 425–428



[31]Surpin M, Raikhel N. Traffic jams affect plant development and signal transduction. Nat Rev Mol Cell Biol, 2004, 5: 100–109



[32]Molendijk A J, Ruperti B, Palme K. Small GTPases in vesicle trafficking. Curr Opin Plant Biol, 2004, 7: 694–700



[33]Vernoud V, Horton A C, Yang Z B, Nielsen E. Analysis of the small GTPase gene superfamily of Arabidopsis. Plant Physiol, 2003, 131: 1191–1208



[34]Peng J L, Ilarslan H, Wurtele E S, Bassham D C. AtRabD2b and AtRabD2c have overlapping functions in pollen development and pollen tube growth. BMC Plant Biol, 2011, 11: 25



[35]Mazel A, Leshem Y, Tiwari B S, Levine A. induction of salt and osmotic stress tolerance by overexpression of an intracellular vesicle trafficking protein AtRab7 (AtRabG3e). Plant Physiol, 2004, 134: 118–128



[36]White P J, Marshall J, Smith J A C. Substrate kinetics of the tonoplast H+-translocating inorganic pyrophosphatase and its activation by free Mg2+. Plant Physiol, 1990, 93: 1063–1070



[37]Parvanova D, Ivanov S, Konstantinova T, Karanovc E, Atanassov A, Tsvetkov T, Alexieva V, Djilianov D. Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress. Plant Physiol Biochem, 2004, 42: 57–63



[38]Boguski M S, McCormick F. Proteins regulating Ras and its relatives. Nature, 1993, 366: 643–654



[39]Takai Y, Sasaki T, Matozaki T: Small GTP-binding proteins. Physiol Rev, 2001, 81: 153–208



[40]Preuss M L, Serna J, Falbel T G, Bednarek S Y, Nielsen E. The Arabidopsis Rab GTPase RabA4b localizes to the tips of growing root hair cells. Plant Cell, 2004, 16: 1589–1603

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