The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase

Kosami, Ken-Ichi; Ohki, Izuru; Nagano, Minoru; Furuita, Kyoko; Sugiki, Toshihiko; Kawano, Yoji; Kawasaki, Tsutomu; Fujiwara, Toshimichi; Nakagawa, Atushi; Shimamoto, Ko; Kojima, Chojiro

doi:info:doi/10.1074/jbc.M114.603282

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タイトル	The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase
著者	Kosami, Ken-Ichi Kosami, Ken-Ichi
	Ohki, Izuru Ohki, Izuru
	Nagano, Minoru Nagano, Minoru
	Furuita, Kyoko Furuita, Kyoko
	Sugiki, Toshihiko Sugiki, Toshihiko
	Kawano, Yoji Kawano, Yoji
	Kawasaki, Tsutomu Kawasaki, Tsutomu
	Fujiwara, Toshimichi Fujiwara, Toshimichi
	Nakagawa, Atushi Nakagawa, Atushi
	Shimamoto, Ko Shimamoto, Ko
	Kojima, Chojiro Kojima, Chojiro
内容	This research was originally published in Journal of Biological Chemistry. Ken-ichi Kosami, Izuru Ohki, Minoru Nagano, Kyoko Furuita, Toshihiko Sugiki, Yoji Kawano, Tsutomu Kawasaki, Toshimichi Fujiwara, Atsushi Nakagawa, Ko Shimamoto and Chojiro Kojima. The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase. Journal of Biological Chemistry. 2014; 289, 28569-28578. © the American Society for Biochemistry and Molecular Biology.
抄録	Rac/Rop proteins are Rho-type small GTPases that act as molecular switches in plants. Recent studies have identified these proteins as key components in many major plant signaling pathways, such as innate immunity, pollen tube growth, and root hair formation. In rice, the Rac/Rop protein OsRac1 plays an important role in regulating the production of reactive oxygen species (ROS) by the NADPH oxidase OsRbohB during innate immunity. However, the molecular mechanism by which OsRac1 regulates OsRbohB remains unknown. Here, we report the crystal structure of OsRac1 complexed with the non-hydrolyzable GTP analog guanosine 5´-(β,γ-imido)triphosphate at 1.9 Å resolution; this represents the first active-form structure of a plant small GTPase. To elucidate theROSproduction in rice cells, structural information was used to design OsRac1 mutants that displayed reduced binding to OsRbohB. Only mutations in the OsRac1 Switch I region showed attenuated interactions with OsRbohB in vitro. In particular, Tyr^39 and Asp^45 substitutions suppressed ROS production in rice cells, indicating that these residues are critical for interaction with and activation of OsRbohB. Structural comparison of active-form OsRac1 with AtRop9 in its GDP-bound inactive form showed a large conformational difference in the vicinity of these residues. Our results provide new insights into the molecular mechanism of the immune response through OsRac1 and the various cellular responses associated with plant Rac/Rop proteins.
公開者	American Society for Biochemistry and Molecular Biology
掲載誌名	Journal of Biological Chemistry
巻	289
号	41
開始ページ	28569
終了ページ	28578
刊行年月	2014-10
ISSN	1083351X
pmid	25128531
NCID	AA1202441X
DOI	info:doi/10.1074/jbc.M114.603282
URL	http://hdl.handle.net/11094/73665
権利情報	© the American Society for Biochemistry and Molecular Biology.
言語	英語
カテゴリ	学術雑誌論文 Journal Article

著者版フラグ	publisher
NII資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
dcmi資源タイプ	text
DCTERMS.bibliographicCitation	Journal of Biological Chemistry.289(41) P.28569-P.28578
DC.title	The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase
DC.creator	Kosami, Ken-Ichi
	Ohki, Izuru
	Nagano, Minoru
	Furuita, Kyoko
	Sugiki, Toshihiko
	Kawano, Yoji
	Kawasaki, Tsutomu
	Fujiwara, Toshimichi
	Nakagawa, Atushi
	Shimamoto, Ko
	Kojima, Chojiro
DC.publisher	American Society for Biochemistry and Molecular Biology
DC.language" scheme="DCTERMS.RFC1766	英語
DCTERMS.issued" scheme="DCTERMS.W3CDTF	2014-10
DC.identifier	info:doi/10.1074/jbc.M114.603282
DC.identifier" scheme="DCTERMS.URI	http://hdl.handle.net/11094/73665
DC.description	This research was originally published in Journal of Biological Chemistry. Ken-ichi Kosami, Izuru Ohki, Minoru Nagano, Kyoko Furuita, Toshihiko Sugiki, Yoji Kawano, Tsutomu Kawasaki, Toshimichi Fujiwara, Atsushi Nakagawa, Ko Shimamoto and Chojiro Kojima. The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase. Journal of Biological Chemistry. 2014; 289, 28569-28578. © the American Society for Biochemistry and Molecular Biology.
DCTERMS.abstract	Rac/Rop proteins are Rho-type small GTPases that act as molecular switches in plants. Recent studies have identified these proteins as key components in many major plant signaling pathways, such as innate immunity, pollen tube growth, and root hair formation. In rice, the Rac/Rop protein OsRac1 plays an important role in regulating the production of reactive oxygen species (ROS) by the NADPH oxidase OsRbohB during innate immunity. However, the molecular mechanism by which OsRac1 regulates OsRbohB remains unknown. Here, we report the crystal structure of OsRac1 complexed with the non-hydrolyzable GTP analog guanosine 5´-(β,γ-imido)triphosphate at 1.9 Å resolution; this represents the first active-form structure of a plant small GTPase. To elucidate theROSproduction in rice cells, structural information was used to design OsRac1 mutants that displayed reduced binding to OsRbohB. Only mutations in the OsRac1 Switch I region showed attenuated interactions with OsRbohB in vitro. In particular, Tyr^39 and Asp^45 substitutions suppressed ROS production in rice cells, indicating that these residues are critical for interaction with and activation of OsRbohB. Structural comparison of active-form OsRac1 with AtRop9 in its GDP-bound inactive form showed a large conformational difference in the vicinity of these residues. Our results provide new insights into the molecular mechanism of the immune response through OsRac1 and the various cellular responses associated with plant Rac/Rop proteins.
DC.rights	© the American Society for Biochemistry and Molecular Biology.
citation_title	The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase
citation_author	Kosami, Ken-Ichi
	Ohki, Izuru
	Nagano, Minoru
	Furuita, Kyoko
	Sugiki, Toshihiko
	Kawano, Yoji
	Kawasaki, Tsutomu
	Fujiwara, Toshimichi
	Nakagawa, Atushi
	Shimamoto, Ko
	Kojima, Chojiro
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	英語
citation_date	2014-10
citation_journal_title	Journal of Biological Chemistry
citation_volume	289
citation_issue	41
citation_firstpage	28569
citation_lastpage	28578
citation_issn	1083351X
citation_doi	info:doi/10.1074/jbc.M114.603282
citation_pmid	25128531
citation_public_url	http://hdl.handle.net/11094/73665

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