Tomato Pto encodes a functional N-myristoylation motif that is required for signal transduction in Nicotiana benthamiana.
Pto kinase of tomato (Lycopersicon esculentum) confers resistance to bacterial speck disease caused by Pseudomonas syringae pv. tomato expressing avrPto or avrPtoB. Pto interacts directly with these type-III secreted effectors, leading to induction of defence responses including the hypersensitive response (HR). Signalling by Pto requires the nucleotide-binding site-leucine-rich repeat (NBS-LRR) protein Prf. Little is known of how Pto is controlled prior to or during stimulation, although kinase activity is required for Avr-dependent activation. Here we demonstrate a role for the N-terminus in signalling by Pto. N-terminal residues outside the kinase domain were required for induction of the HR in Nicotiana benthamiana. The N-terminus also contributed to both AvrPto-binding and phosphorylation abilities. Pto residues 1-10 comprise a consensus motif for covalent attachment of myristate, a hydrophobic 14-carbon saturated fatty acid, to the Gly-2 residue. Several lines of evidence indicate that this motif is important for Pto function. A heterologous N-myristoylation motif complemented N-terminal deletion mutants of Pto for Prf-dependent signalling. Signalling by wild-type and mutant forms of Pto was strictly dependent on the Gly-2 residue. The N-myristoylation motif of Pto complemented the cognate motif of AvrPto for avirulence function and membrane association. Furthermore, Pto was myristoylated in vivo dependent on the presence of Gly-2. The subcellular localization of Pto was independent of N-myristoylation, indicating that N-myristoylation is required for some function other than membrane affinity. Consistent with this idea, AvrPtoB was also found to be a soluble protein. The data indicate an important role(s) for the myristoylated N-terminus in Pto signalling.
de Vries
Jeroen S
JS
The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK.
Andriotis
Vasilios M E
VM
Wu
Ai-Jiuan
AJ
Rathjen
John P
JP
D016960:Agrobacterium tumefaciens / Q000235:genetics; D000596:Amino Acids / Q000378:metabolism; D018551:Solanum lycopersicum / Q000235:genetics*; D019814:Myristic Acid / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D015398:Signal Transduction; D014026:Nicotiana / Q000378:metabolism*
D000596:Amino Acids; D010940:Plant Proteins; D019814:Myristic Acid; C084354:Pto protein, Lycopersicon; D017346:Protein Serine-Threonine Kinases