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:https://doi.org/10.1074/jbc.M114.603282

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

Kosami, Ken-ichi; Ohki, Izuru; Nagano, Minoru et al.

Journal of Biological Chemistry, 2014, 289(41), 28569-28578

Number of Access:1,336（2025-09-29 10:03 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/73665

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Title	The crystal structure of the plant small GTPase OsRac1 reveals its mode of binding to NADPH oxidase
Creator	CreatorName Kosami, Ken-ichi
	NRID 1000080418574 1000080418574 CreatorName Ohki, Izuru
	NRID 1000080598251 1000080598251 CreatorName Nagano, Minoru
	NRID 1000030727665 1000030727665 researchmap permalink k-furuita k-furuita CreatorName Furuita, Kyoko
	NRID 1000070635698 1000070635698 CreatorName Sugiki, Toshihiko
	NRID 1000000406175 1000000406175 CreatorName Kawano, Yoji
	NRID 1000090283936 1000090283936 CreatorName Kawasaki, Tsutomu
	NRID 1000020242381 1000020242381 researchmap permalink read0009115 read0009115 Scopus Author ID 7403542603 7403542603 CreatorName Fujiwara, Toshimichi
	NRID 1000020188890 1000020188890 ORCID 0000-0002-1700-7861 0000-0002-1700-7861 researchmap permalink atsushi_nakagawa atsushi_nakagawa Scopus Author ID 7202934909 7202934909 CreatorName Nakagawa, Atushi
	NRID 1000010263427 1000010263427 CreatorName Shimamoto, Ko
	NRID 1000050333563 1000050333563 ORCID 0000-0003-2723-8249 0000-0003-2723-8249 researchmap permalink read0072023 read0072023 CreatorName Kojima, Chojiro
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.
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.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	289(41)
Page	28569-28578
Date of Issued	2014-10
Language	English
Handle URL	https://hdl.handle.net/11094/73665
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M114.603282
Relation.isIdenticalTo	PMID 25128531
Access Rights	open access
CopyRight	© the American Society for Biochemistry and Molecular Biology.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Journal of Biological Chemistry. 2014, 289(41), p. 28569-28578
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	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2014-10
DC.identifier	https://doi.org/10.1074/jbc.M114.603282
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.
DC.format	73665
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	English
citation_date	2014-10
citation_journal_title	Journal of Biological Chemistry
citation_volume	289
citation_issue	41
citation_firstpage	28569
citation_lastpage	28578
citation_doi	https://doi.org/10.1074/jbc.M114.603282
citation_pmid	25128531
citation_public_url	https://hdl.handle.net/11094/73665

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

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