Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension

Hann Ling Wong, Reinhard Pinontoan, Kokoro Hayashi, Ryo Tabata, Takashi Yaeno, Kana Hasegawa, Chojiro Kojima, Hirofumi Yoshioka, Koh Iba, Tsutomu Kawasaki, Ko Shimamoto

Research output: Contribution to journalArticle

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Abstract

Reactive oxygen species (ROS) produced by NADPH oxidase play critical roles in various cellular activities, including plant innate immunity response. In contrast with the large multiprotein NADPH oxidase complex of phagocytes, in plants, only the homologs of the catalytic subunit gp91phox and the cytosolic regulator small GTPase Rac are found. Plant homologs of the gp91 phox subunit are known as Rboh (for respiratory burst oxidase homolog). Although numerous Rboh have been isolated in plants, the regulation of enzymatic activity remains unknown. All rboh genes identified to date possess a conserved N-terminal extension that contains two Ca2+ binding EF-hand motifs. Previously, we ascertained that a small GTPase Rac (Os Rac1) enhanced pathogen-associated molecular pattern-induced ROS production and resistance to pathogens in rice (Oryza sativa). In this study, using yeast two-hybrid assay, we found that interaction between Rac GTPases and the N-terminal extension is ubiquitous and that a substantial part of the N-terminal region of Rboh, including the two EF-hand motifs, is required for the interaction. The direct Rac-Rboh interaction was supported by further studies using in vitro pull-down assay, a nuclear magnetic resonance titration experiment, and in vivo fluorescence resonance energy transfer (FRET) microscopy. The FRET analysis also suggests that cytosolic Ca2+ concentration may regulate Rac-Rboh interaction in a dynamic manner. Furthermore, transient coexpression of Os Rac1 and rbohB enhanced ROS production in Nicotiana benthamiana, suggesting that direct Rac-Rboh interaction may activate NADPH oxidase activity in plants. Taken together, the results suggest that cytosolic Ca2+ concentration may modulate NADPH oxidase activity by regulating the interaction between Rac GTPase and Rboh.

Original languageEnglish
Pages (from-to)4022-4034
Number of pages13
JournalPlant Cell
Volume19
Issue number12
DOIs
Publication statusPublished - Dec 1 2007

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NADPH Oxidase
GTP Phosphohydrolases
guanosinetriphosphatase
EF hand motifs
rice
reactive oxygen species
energy transfer
calcium
EF Hand Motifs
Reactive Oxygen Species
Fluorescence Resonance Energy Transfer
Monomeric GTP-Binding Proteins
fluorescence
two hybrid system techniques
Nicotiana benthamiana
pathogens
phagocytes
Plant Immunity
protein subunits
titration

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology

Cite this

Wong, H. L., Pinontoan, R., Hayashi, K., Tabata, R., Yaeno, T., Hasegawa, K., ... Shimamoto, K. (2007). Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension. Plant Cell, 19(12), 4022-4034. https://doi.org/10.1105/tpc.107.055624

Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension. / Wong, Hann Ling; Pinontoan, Reinhard; Hayashi, Kokoro; Tabata, Ryo; Yaeno, Takashi; Hasegawa, Kana; Kojima, Chojiro; Yoshioka, Hirofumi; Iba, Koh; Kawasaki, Tsutomu; Shimamoto, Ko.

In: Plant Cell, Vol. 19, No. 12, 01.12.2007, p. 4022-4034.

Research output: Contribution to journalArticle

Wong, HL, Pinontoan, R, Hayashi, K, Tabata, R, Yaeno, T, Hasegawa, K, Kojima, C, Yoshioka, H, Iba, K, Kawasaki, T & Shimamoto, K 2007, 'Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension', Plant Cell, vol. 19, no. 12, pp. 4022-4034. https://doi.org/10.1105/tpc.107.055624
Wong HL, Pinontoan R, Hayashi K, Tabata R, Yaeno T, Hasegawa K et al. Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension. Plant Cell. 2007 Dec 1;19(12):4022-4034. https://doi.org/10.1105/tpc.107.055624
Wong, Hann Ling ; Pinontoan, Reinhard ; Hayashi, Kokoro ; Tabata, Ryo ; Yaeno, Takashi ; Hasegawa, Kana ; Kojima, Chojiro ; Yoshioka, Hirofumi ; Iba, Koh ; Kawasaki, Tsutomu ; Shimamoto, Ko. / Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension. In: Plant Cell. 2007 ; Vol. 19, No. 12. pp. 4022-4034.
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AU - Yaeno, Takashi

AU - Hasegawa, Kana

AU - Kojima, Chojiro

AU - Yoshioka, Hirofumi

AU - Iba, Koh

AU - Kawasaki, Tsutomu

AU - Shimamoto, Ko

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