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PLANT CELL 16 (10): 2809-2821

Copyright © 2004 by the American Society of Plant Physiologists.

A Patch of Surface-Exposed Residues Mediates Negative Regulation of Immune Signaling by Tomato Pto Kinase{boxw}

Ai-Jiuan Wua, Vasilios M.E. Andriotisa, Marcus C. Durrantb, and John P. Rathjena,1

a Sainsbury Laboratory, Norwich Research Park, Norwich, Norfolk, NR4 7UH, United Kingdom
b Computational Biology Group, John Innes Centre, Norwich, Norfolk, NR4 7UH, United Kingdom

1 To whom correspondence should be addressed. E-mail john.rathjen{at}sainsbury-laboratory.ac.uk; fax 44-1603-450-011.

Abstract: Tomato (Lycopersicon esculentum) Pto kinase specifically recognizes the Pseudomonas effector proteins AvrPto and AvrPtoB, leading to induction of defense responses and hypersensitive cell death. Structural modeling of Pto combined with site-directed mutagenesis identified a patch of surface-exposed residues required for native regulation of signaling. Mutations in this area resulted in constitutive gain-of-function (CGF) forms of Pto that activated AvrPto-independent cell death via the cognate signaling pathway. The patch overlaps the peptide binding region of the kinase catalytic cleft and is part of a broader region required for interaction with bacterial effectors. We propose that the negative regulatory patch is normally occupied by a peptide that represses Pto signaling. Furthermore, we found that Pto kinase activity was required for Avr-dependent activation but dispensable for signaling by CGF forms of Pto. This suggests that Pto signals by a conformational change rather than phosphorylation of downstream substrates in the defense signaling pathway.

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