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GrainGenes Reference Report: PLA-210-143

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Reference	PLA-210-143
Title	Characterization of two fungal-elicitor-induced rice cDNAs encoding functional homologues of the rab-specific GDP-dissociation inhibitor
Journal	Planta
Year	1999
Volume	210
Pages	143-149
Author	Kim WY [ Show all 10 ]
Abstract	By using the mRNA differential display approach to isolate defense signaling genes active at the early stage of fungal infection two cDNA fragments with high sequence homology to rab-specific GDP-dissociation inhibitors (GDIs) were identified in rice (Oryza sativa L.) suspension cells. Using polymerase-chain-reaction products as probes, two full-length cDNA clones were isolated from a cDNA library of fungal-elicitor-treated rice, and designated as OsGDII and OsGDI2. The deduced amino acid sequences of the isolated cDNAs exhibited substantial homology to Arabidopsis rab-GDIs. Northern analysis revealed that transcripts detected with the 3'-gene-specific DNA probes accumulated to high levels within 30 min after treatment with a fungal elicitor derived from Magnaporthe grisea. The functionality of the OsGDIs was demonstrated by their ability to rescue the Sec19 mutant of Saccharomyces cerevisiae which is defective in vesicle transport. The proteins, expressed in Escherichia coli, cross-reacted with a polyclonal antibody prepared against bovine rab-GDI. Like bovine rab-GDI, the OsGDI proteins efficiently dissociated rab3A from bovine synaptic membranes. Using the two-hybrid system, it was shown that the OsGDIs specifically interact with the small GTP-binding proteins belonging to the rab subfamily. The specific interaction was also demonstrated in vitro by glutathione S-transferase resin pull-down assay
Keyword	[ Hide all but 1 of 51 ] amino acid amino acid sequence antibody arabidopsis thaliana assay cdna cdna clone cell suspensions clone complementary dna complementation defense differential display dna dna probes elicitor escherichia coli functionality fungal fungal elicitor gene gene expression glutathione glutathione-s-transferase gtp binding gtp binding protein guanosine diphosphate homology in vitro infection inhibitor library magnaporthe grisea membrane messenger rna mrna mutant oligosaccharides oryza sativa polymerase chain reaction probes rab recombinant proteins rescue rice saccharomyces cerevisiae sequence suspension transcript transport two-hybrid system

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