A Plant Homolog of the Neutrophil NADPH Oxidase gp91^phox Subunit Gene Encodes a Plasma Membrane Protein with Ca²⁺ Binding Motifs

Correspondence to: Chris Lamb, lamb{at}salk.edu (E-mail), 619-558-6379 (fax).

Rapid generation of O₂- and H₂O₂, which is reminiscent of the oxidative burst in neutrophils, is a central component of the resistance response of plants to pathogen challenge. Here, we report that the Arabidopsis rbohA (for respiratory burst oxidase homolog A) gene encodes a putative 108-kD protein, with a C-terminal region that shows pronounced similarity to the 69-kD apoprotein of the gp91^phox subunit of the neutrophil respiratory burst NADPH oxidase. The RbohA protein has a large hydrophilic N-terminal domain that is not present in gp91^phox. This domain contains two Ca²⁺ binding EF hand motifs and has extended similarity to the human RanGTPase-activating protein 1. rbohA, which is a member of a divergent gene family, generates transcripts of 3.6 and 4.0 kb that differ only in their polyadenylation sites. rbohA transcripts are most abundant in roots, with weaker expression in aerial organs and seedlings. Antibodies raised against a peptide near the RbohA C terminus detected a 105-kD protein that, unlike gp91^phox, does not appear to be highly glycosylated. Cell fractionation, two-phase partitioning, and detergent extraction indicate that RbohA is an intrinsic plasma membrane protein. We propose that plants have a plasma membrane enzyme similar to the neutrophil NADPH oxidase but with novel potential regulatory mechanisms for Ca²⁺ and G protein stimulation of O₂- and H₂O₂ production at the cell surface.

This article has been cited by other articles:

				R. Errakhi, A. Dauphin, P. Meimoun, A. Lehner, D. Reboutier, P. Vatsa, J. Briand, K. Madiona, J. P. Rona, M. Barakate, et al. An early Ca2+ influx is a prerequisite to thaxtomin A-induced cell death in Arabidopsis thaliana cells J. Exp. Bot., November 17, 2008; (2008) ern267v1. [Abstract] [Full Text] [PDF]

				D. Zhang, D. Wengier, B. Shuai, C.-P. Gui, J. Muschietti, S. McCormick, and W.-H. Tang The Pollen Receptor Kinase LePRK2 Mediates Growth-Promoting Signals and Positively Regulates Pollen Germination and Tube Growth Plant Physiology, November 1, 2008; 148(3): 1368 - 1379. [Abstract] [Full Text] [PDF]

				A. Savidor, R. S. Donahoo, O. Hurtado-Gonzales, M. L. Land, M. B. Shah, K. H. Lamour, and W. H. McDonald Cross-species Global Proteomics Reveals Conserved and Unique Processes in Phytophthora sojae and Phytophthora ramorum Mol. Cell. Proteomics, August 1, 2008; 7(8): 1501 - 1516. [Abstract] [Full Text] [PDF]

				Y. Lee, G. Bak, Y. Choi, W.-I Chuang, H.-T. Cho, and Y. Lee Roles of Phosphatidylinositol 3-Kinase in Root Hair Growth Plant Physiology, June 1, 2008; 147(2): 624 - 635. [Abstract] [Full Text] [PDF]

				P. Schopfer, E. Heyno, F. Drepper, and A. Krieger-Liszkay Naphthoquinone-Dependent Generation of Superoxide Radicals by Quinone Reductase Isolated from the Plasma Membrane of Soybean Plant Physiology, June 1, 2008; 147(2): 864 - 878. [Abstract] [Full Text] [PDF]

				S. Asai, K. Ohta, and H. Yoshioka MAPK Signaling Regulates Nitric Oxide and NADPH Oxidase-Dependent Oxidative Bursts in Nicotiana benthamiana PLANT CELL, May 1, 2008; 20(5): 1390 - 1406. [Abstract] [Full Text] [PDF]

				Y. Ogasawara, H. Kaya, G. Hiraoka, F. Yumoto, S. Kimura, Y. Kadota, H. Hishinuma, E. Senzaki, S. Yamagoe, K. Nagata, et al. Synergistic Activation of the Arabidopsis NADPH Oxidase AtrbohD by Ca2+ and Phosphorylation J. Biol. Chem., April 4, 2008; 283(14): 8885 - 8892. [Abstract] [Full Text] [PDF]

				S. Takeda, C. Gapper, H. Kaya, E. Bell, K. Kuchitsu, and L. Dolan Local Positive Feedback Regulation Determines Cell Shape in Root Hair Cells Science, February 29, 2008; 319(5867): 1241 - 1244. [Abstract] [Full Text] [PDF]

				F. Weinberger Pathogen-Induced Defense and Innate Immunity in Macroalgae Biol. Bull., December 1, 2007; 213(3): 290 - 302. [Abstract] [Full Text] [PDF]

				H. L. Wong, R. Pinontoan, K. Hayashi, R. Tabata, T. Yaeno, K. Hasegawa, C. Kojima, H. Yoshioka, K. Iba, T. Kawasaki, et al. Regulation of Rice NADPH Oxidase by Binding of Rac GTPase to Its N-Terminal Extension PLANT CELL, December 1, 2007; 19(12): 4022 - 4034. [Abstract] [Full Text] [PDF]

				H. W. Choi, Y. J. Kim, S. C. Lee, J. K. Hong, and B. K. Hwang Hydrogen Peroxide Generation by the Pepper Extracellular Peroxidase CaPO2 Activates Local and Systemic Cell Death and Defense Response to Bacterial Pathogens Plant Physiology, November 1, 2007; 145(3): 890 - 904. [Abstract] [Full Text] [PDF]

				J. Ding, Y. Sun, C. L. Xiao, K. Shi, Y. H. Zhou, and J. Q. Yu Physiological basis of different allelopathic reactions of cucumber and figleaf gourd plants to cinnamic acid J. Exp. Bot., October 26, 2007; (2007) erm227v1. [Abstract] [Full Text] [PDF]

				M. J. Egan, Z.-Y. Wang, M. A. Jones, N. Smirnoff, and N. J. Talbot Generation of reactive oxygen species by fungal NADPH oxidases is required for rice blast disease PNAS, July 10, 2007; 104(28): 11772 - 11777. [Abstract] [Full Text] [PDF]

				S. Peleg-Grossman, H. Volpin, and A. Levine Root hair curling and Rhizobium infection in Medicago truncatula are mediated by phosphatidylinositide-regulated endocytosis and reactive oxygen species J. Exp. Bot., May 1, 2007; 58(7): 1637 - 1649. [Abstract] [Full Text] [PDF]

				M. A. Jones, M. J. Raymond, Z. Yang, and N. Smirnoff NADPH oxidase-dependent reactive oxygen species formation required for root hair growth depends on ROP GTPase J. Exp. Bot., April 1, 2007; 58(6): 1261 - 1270. [Abstract] [Full Text] [PDF]

				C. Ashtamker, V. Kiss, M. Sagi, O. Davydov, and R. Fluhr Diverse Subcellular Locations of Cryptogein-Induced Reactive Oxygen Species Production in Tobacco Bright Yellow-2 Cells Plant Physiology, April 1, 2007; 143(4): 1817 - 1826. [Abstract] [Full Text] [PDF]

				M. Kobayashi, I. Ohura, K. Kawakita, N. Yokota, M. Fujiwara, K. Shimamoto, N. Doke, and H. Yoshioka Calcium-Dependent Protein Kinases Regulate the Production of Reactive Oxygen Species by Potato NADPH Oxidase PLANT CELL, March 1, 2007; 19(3): 1065 - 1080. [Abstract] [Full Text] [PDF]

				C. Carter, R. Healy, N. M. O'Tool, S.M. S. Naqvi, G. Ren, S. Park, G. A. Beattie, H. T. Horner, and R. W. Thornburg Tobacco Nectaries Express a Novel NADPH Oxidase Implicated in the Defense of Floral Reproductive Tissues against Microorganisms Plant Physiology, January 1, 2007; 143(1): 389 - 399. [Abstract] [Full Text] [PDF]

				M. Trujillo, L. Altschmied, P. Schweizer, K.-H. Kogel, and R. Huckelhoven Respiratory Burst Oxidase Homologue A of barley contributes to penetration by the powdery mildew fungus Blumeria graminis f. sp. hordei J. Exp. Bot., November 1, 2006; 57(14): 3781 - 3791. [Abstract] [Full Text] [PDF]

				H. Yoda, Y. Hiroi, and H. Sano Polyamine Oxidase Is One of the Key Elements for Oxidative Burst to Induce Programmed Cell Death in Tobacco Cultured Cells Plant Physiology, September 1, 2006; 142(1): 193 - 206. [Abstract] [Full Text] [PDF]

				K. Kojo, T. Yaeno, K. Kusumi, H. Matsumura, S. Fujisawa, R. Terauchi, and K. Iba Regulatory Mechanisms of ROI Generation are Affected by Rice spl Mutations Plant Cell Physiol., August 1, 2006; 47(8): 1035 - 1044. [Abstract] [Full Text] [PDF]

				C. Gapper and L. Dolan Control of Plant Development by Reactive Oxygen Species Plant Physiology, June 1, 2006; 141(2): 341 - 345. [Full Text] [PDF]

				D. L. BOURANIS, S. N. CHORIANOPOULOU, C. KOLLIAS, P. MANIOU, V. E. PROTONOTARIOS, V. F. SIYIANNIS, and M. J. HAWKESFORD Dynamics of Aerenchyma Distribution in the Cortex of Sulfate-deprived Adventitious Roots of Maize Ann. Bot., May 1, 2006; 97(5): 695 - 704. [Abstract] [Full Text] [PDF]

				R. J Carol and L. Dolan The role of reactive oxygen species in cell growth: lessons from root hairs J. Exp. Bot., May 1, 2006; 57(8): 1829 - 1834. [Abstract] [Full Text] [PDF]

				D. R Davies, L. V Bindschedler, T. S Strickland, and G P. Bolwell Production of reactive oxygen species in Arabidopsis thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum: implications for basal resistance J. Exp. Bot., May 1, 2006; 57(8): 1817 - 1827. [Abstract] [Full Text] [PDF]

				C. J. Song, I. Steinebrunner, X. Wang, S. C. Stout, and S. J. Roux Extracellular ATP Induces the Accumulation of Superoxide via NADPH Oxidases in Arabidopsis Plant Physiology, April 1, 2006; 140(4): 1222 - 1232. [Abstract] [Full Text] [PDF]

				M. Kobayashi, K. Kawakita, M. Maeshima, N. Doke, and H. Yoshioka Subcellular localization of Strboh proteins and NADPH-dependent O2--generating activity in potato tuber tissues J. Exp. Bot., March 1, 2006; 57(6): 1373 - 1379. [Abstract] [Full Text] [PDF]

				C. Yamamizo, K. Kuchimura, A. Kobayashi, S. Katou, K. Kawakita, J. D.G. Jones, N. Doke, and H. Yoshioka Rewiring Mitogen-Activated Protein Kinase Cascade by Positive Feedback Confers Potato Blight Resistance Plant Physiology, February 1, 2006; 140(2): 681 - 692. [Abstract] [Full Text] [PDF]

				C. Delis, M. Dimou, E. Flemetakis, G. Aivalakis, and P. Katinakis A root- and hypocotyl-specific gene coding for copper-containing amine oxidase is related to cell expansion in soybean seedlings J. Exp. Bot., January 1, 2006; 57(1): 101 - 111. [Abstract] [Full Text] [PDF]

				T. Kawahara, D. Ritsick, G. Cheng, and J. D. Lambeth Point Mutations in the Proline-rich Region of p22phox Are Dominant Inhibitors of Nox1- and Nox2-dependent Reactive Oxygen Generation J. Biol. Chem., September 9, 2005; 280(36): 31859 - 31869. [Abstract] [Full Text] [PDF]

				J. BAILEY-SERRES and R. CHANG Sensing and Signalling in Response to Oxygen Deprivation in Plants and Other Organisms Ann. Bot., September 1, 2005; 96(4): 507 - 518. [Abstract] [Full Text] [PDF]

				T. Brembu, P. Winge, and A. M. Bones The small GTPase AtRAC2/ROP7 is specifically expressed during late stages of xylem differentiation in Arabidopsis J. Exp. Bot., September 1, 2005; 56(419): 2465 - 2476. [Abstract] [Full Text] [PDF]

				Y.-L. Chen, R. Huang, Y.-M. Xiao, P. Lu, J. Chen, and X.-C. Wang Extracellular Calmodulin-Induced Stomatal Closure Is Mediated by Heterotrimeric G Protein and H2O2 Plant Physiology, December 1, 2004; 136(4): 4096 - 4103. [Abstract] [Full Text] [PDF]

				T. Nagata, S. Iizumi, K. Satoh, H. Ooka, J. Kawai, P. Carninci, Y. Hayashizaki, Y. Otomo, K. Murakami, K. Matsubara, et al. Comparative Analysis of Plant and Animal Calcium Signal Transduction Element Using Plant Full-Length cDNA Data Mol. Biol. Evol., October 1, 2004; 21(10): 1855 - 1870. [Abstract] [Full Text] [PDF]

				K. H. Nam and J. Li The Arabidopsis Transthyretin-Like Protein Is a Potential Substrate of BRASSINOSTEROID-INSENSITIVE 1 PLANT CELL, September 1, 2004; 16(9): 2406 - 2417. [Abstract] [Full Text] [PDF]

				S. Lorrain, B. Lin, M. C. Auriac, T. Kroj, P. Saindrenan, M. Nicole, C. Balague, and D. Roby VASCULAR ASSOCIATED DEATH1, a Novel GRAM Domain-Containing Protein, Is a Regulator of Cell Death and Defense Responses in Vascular Tissues PLANT CELL, August 1, 2004; 16(8): 2217 - 2232. [Abstract] [Full Text] [PDF]

				I. C. Mori and J. I. Schroeder Reactive Oxygen Species Activation of Plant Ca2+ Channels. A Signaling Mechanism in Polar Growth, Hormone Transduction, Stress Signaling, and Hypothetically Mechanotransduction Plant Physiology, June 1, 2004; 135(2): 702 - 708. [Full Text] [PDF]

				M. Sagi, O. Davydov, S. Orazova, Z. Yesbergenova, R. Ophir, J. W. Stratmann, and R. Fluhr Plant Respiratory Burst Oxidase Homologs Impinge on Wound Responsiveness and Development in Lycopersicon esculentum PLANT CELL, March 1, 2004; 16(3): 616 - 628. [Abstract] [Full Text] [PDF]

				R. Desikan, M.-K. Cheung, J. Bright, D. Henson, J. T. Hancock, and S. J. Neill ABA, hydrogen peroxide and nitric oxide signalling in stomatal guard cells J. Exp. Bot., January 2, 2004; 55(395): 205 - 212. [Abstract] [Full Text] [PDF]

				J. Park, Y. Gu, Y. Lee, Z. Yang, and Y. Lee Phosphatidic Acid Induces Leaf Cell Death in Arabidopsis by Activating the Rho-Related Small G Protein GTPase-Mediated Pathway of Reactive Oxygen Species Generation Plant Physiology, January 1, 2004; 134(1): 129 - 136. [Abstract] [Full Text] [PDF]

				A. Ranieri, A. Castagna, J. Pacini, B. Baldan, A. Mensuali Sodi, and G. F. Soldatini Early production and scavenging of hydrogen peroxide in the apoplast of sunflower plants exposed to ozone J. Exp. Bot., November 1, 2003; 54(392): 2529 - 2540. [Abstract] [Full Text] [PDF]

				H. Yoda, Y. Yamaguchi, and H. Sano Induction of Hypersensitive Cell Death by Hydrogen Peroxide Produced through Polyamine Degradation in Tobacco Plants Plant Physiology, August 1, 2003; 132(4): 1973 - 1981. [Abstract] [Full Text] [PDF]

				H. Yoshioka, N. Numata, K. Nakajima, S. Katou, K. Kawakita, O. Rowland, J. D. G. Jones, and N. Doke Nicotiana benthamiana gp91phox Homologs NbrbohA and NbrbohB Participate in H2O2 Accumulation and Resistance to Phytophthora infestans PLANT CELL, March 1, 2003; 15(3): 706 - 718. [Abstract] [Full Text] [PDF]

				J. Zhao and K. Sakai Multiple signalling pathways mediate fungal elicitor-induced {beta}-thujaplicin biosynthesis in Cupressus lusitanica cell cultures J. Exp. Bot., February 1, 2003; 54(383): 647 - 656. [Abstract] [Full Text] [PDF]

				E. Olmos, J. R. Martinez-Solano, A. Piqueras, and E. Hellin Early steps in the oxidative burst induced by cadmium in cultured tobacco cells (BY-2 line) J. Exp. Bot., January 2, 2003; 54(381): 291 - 301. [Abstract] [Full Text] [PDF]

				S. Bourque, R. Lemoine, A. Sequeira-Legrand, L. Fayolle, S. Delrot, and A. Pugin The Elicitor Cryptogein Blocks Glucose Transport in Tobacco Cells Plant Physiology, December 1, 2002; 130(4): 2177 - 2187. [Abstract] [Full Text] [PDF]

				B. Klusener, J. J. Young, Y. Murata, G. J. Allen, I. C. Mori, V. Hugouvieux, and J. I. Schroeder Convergence of Calcium Signaling Pathways of Pathogenic Elicitors and Abscisic Acid in Arabidopsis Guard Cells Plant Physiology, December 1, 2002; 130(4): 2152 - 2163. [Abstract] [Full Text] [PDF]

				J. Zhao, P. Peng, R. J. Schmitz, A. D. Decker, F. E. Tax, and J. Li Two Putative BIN2 Substrates Are Nuclear Components of Brassinosteroid Signaling Plant Physiology, November 1, 2002; 130(3): 1221 - 1229. [Abstract] [Full Text] [PDF]

				J. C. Moreno, H. Bikker, M. J.E. Kempers, A.S. P. van Trotsenburg, F. Baas, J. J.M. de Vijlder, T. Vulsma, and C. Ris-Stalpers Inactivating Mutations in the Gene for Thyroid Oxidase 2 (THOX2) and Congenital Hypothyroidism N. Engl. J. Med., July 11, 2002; 347(2): 95 - 102. [Abstract] [Full Text] [PDF]

				S.-H. Cheng, M. R. Willmann, H.-C. Chen, and J. Sheen Calcium Signaling through Protein Kinases. The Arabidopsis Calcium-Dependent Protein Kinase Gene Family Plant Physiology, June 1, 2002; 129(2): 469 - 485. [Abstract] [Full Text] [PDF]

				S. J. Neill, R. Desikan, A. Clarke, R. D. Hurst, and J. T. Hancock Hydrogen peroxide and nitric oxide as signalling molecules in plants J. Exp. Bot., May 15, 2002; 53(372): 1237 - 1247. [Abstract] [Full Text] [PDF]

				G. P. Bolwell, L. V. Bindschedler, K. A. Blee, V. S. Butt, D. R. Davies, S. L. Gardner, C. Gerrish, and F. Minibayeva The apoplastic oxidative burst in response to biotic stress in plants: a three-component system J. Exp. Bot., May 15, 2002; 53(372): 1367 - 1376. [Abstract] [Full Text] [PDF]

				T. Yang and B. W. Poovaiah Hydrogen peroxide homeostasis: Activation of plant catalase by calcium/calmodulin PNAS, March 19, 2002; 99(6): 4097 - 4102. [Abstract] [Full Text] [PDF]

				L. Costet, S. Dorey, B. Fritig, and S. Kauffmann A Pharmacological Approach to Test the Diffusible Signal Activity of Reactive Oxygen Intermediates in Elicitor-Treated Tobacco Leaves Plant Cell Physiol., January 1, 2002; 43(1): 91 - 98. [Abstract] [Full Text] [PDF]

				M. A. Torres, J. L. Dangl, and J. D. G. Jones Arabidopsis gp91phox homologues AtrbohD and AtrbohF are required for accumulation of reactive oxygen intermediates in the plant defense response PNAS, December 21, 2001; (2001) 12452499. [Abstract] [Full Text] [PDF]

				Y. Murata, Z.-M. Pei, I. C. Mori, and J. Schroeder Abscisic Acid Activation of Plasma Membrane Ca2+ Channels in Guard Cells Requires Cytosolic NAD(P)H and Is Differentially Disrupted Upstream and Downstream of Reactive Oxygen Species Production in abi1-1 and abi2-1 Protein Phosphatase 2C Mutants PLANT CELL, November 1, 2001; 13(11): 2513 - 2523. [Abstract] [Full Text] [PDF]

				R. Desikan, S. A.-H.-Mackerness, J. T. Hancock, and S. J. Neill Regulation of the Arabidopsis Transcriptome by Oxidative Stress Plant Physiology, September 1, 2001; 127(1): 159 - 172. [Abstract] [Full Text] [PDF]

				Y. Sang, D. Cui, and X. Wang Phospholipase D and Phosphatidic Acid-Mediated Generation of Superoxide in Arabidopsis Plant Physiology, August 1, 2001; 126(4): 1449 - 1458. [Abstract] [Full Text] [PDF]