Palmitoylation-dependent membrane localization of the rice resistance protein pit is critical for the activation of the small GTPase OsRac1.
Nucleotide binding domain and leucine-rich repeat (NLR)-containing family proteins function as intracellular immune sensors in both plants and animals. In plants, the downstream components activated by NLR family proteins and the immune response mechanisms induced by these downstream molecules are largely unknown. We have previously found that the small GTPase OsRac1, which acts as a molecular switch in rice immunity, is activated by Pit, an NLR-type resistance (R) protein to rice blast fungus, and this activation plays critical roles in Pit-mediated immunity. However, the sites and mechanisms of activation of Pit in vivo remain unknown. To clarify the mechanisms involved in the localization of Pit, we searched for consensus sequences in Pit that specify membrane localization and found a pair of potential palmitoylation sites in the N-terminal coiled-coil region. Although wild-type Pit was localized mainly to the plasma membrane, this membrane localization was compromised in a palmitoylation-deficient mutant of Pit. The palmitoylation-deficient Pit displayed significantly lower affinity for OsRac1 on the plasma membrane, thereby resulting in failures of the Pit-mediated cell death, the production of reactive oxygen species, and disease resistance to rice blast fungus. These results indicate that palmitoylation-dependent membrane localization of Pit is required for the interaction with and the activation of OsRac1 and that OsRac1 activation by Pit is vital for Pit-mediated disease resistance to rice blast fungus.
From the Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan and y-kawano@bs.naist.jp.
Yoji
Y
Kawano
From the Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan and.
Tadashi
T
Fujiwara
From the Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan and.
Ai
A
Yao
From the Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan and.
Yusuke
Y
Housen
National Agricultural Research Center, National Agriculture and Food Research Organization, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-8666, Japan.
Keiko
K
Hayashi
From the Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan and.
Ko
K
Shimamoto
D002462:Cell Membrane / Q000378:metabolism*; D060467:Disease Resistance; D004789:Enzyme Activation; D054878:Lipoylation; D012275:Oryza / Q000166:cytology* / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D021381:Protein Transport; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
D010940:Plant Proteins; D020830:rac1 GTP-Binding Protein
Plant Defense; Plant Pathology; Plasma Membrane; Protein Palmitoylation; Small GTPase