The tomato kinome and the tomato kinase library ORFeome: novel resources for the study of kinases and signal transduction in tomato and solanaceae species

Mol Plant Microbe Interact. 2014 Jan;27(1):7-17. doi: 10.1094/MPMI-08-13-0218-TA.

Abstract

Protein kinase-driven phosphorylation constitutes the core of cellular signaling. Kinase components of signal transduction pathways are often targeted for inactivation by pathogens. The study of kinases and immune signal transduction in the model crop tomato (Solanum lycopersicum) would benefit from the availability of community-wide resources for large scale and systems-level experimentation. Here, we defined the tomato kinome and performed a comprehensive comparative analysis of the tomato kinome and 15 other plant species. We constructed a tomato kinase library (TOKN 1.0) of over 300 full-length open reading frames (ORF) cloned into a recombination-based vector. We developed a high-throughput pipeline to isolate and transform tomato protoplasts. A subset of the TOKN 1.0 library kinases were expressed in planta, were purified, and were used to generate a functional tomato protein microarray. All resources created were utilized to test known and novel associations between tomato kinases and Pseudomonas syringae DC3000 effectors in a large-scale format. Bsk7 was identified as a component of the plant immune response and a candidate effector target. These resources will enable comprehensive investigations of signaling pathways and host-pathogen interactions in tomato and other Solanaceae spp.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Computational Biology
  • Gene Library
  • Genetic Complementation Test
  • Host-Pathogen Interactions
  • Luciferases
  • Open Reading Frames
  • Plant Diseases / immunology*
  • Plant Diseases / microbiology
  • Plant Immunity
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Protein Array Analysis
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Protoplasts
  • Pseudomonas syringae / genetics
  • Pseudomonas syringae / metabolism*
  • Signal Transduction*
  • Solanaceae / enzymology
  • Solanaceae / genetics
  • Solanaceae / immunology
  • Solanaceae / physiology*
  • Solanum lycopersicum / enzymology
  • Solanum lycopersicum / genetics
  • Solanum lycopersicum / immunology
  • Solanum lycopersicum / physiology*

Substances

  • Bacterial Proteins
  • Plant Proteins
  • Luciferases
  • Protein Kinases