A Protein Interaction Database For Rice Protein Kinases

National Science Foundation DBI-0217312


                                 

Project materials



| PlantsP | Top | Abstract | Objectives | Deliverables | Targets | Private materials |

Project Abstract

The cereals, including rice, maize, wheat, barley, sorghum, and the millets, are the most important group of plants in agriculture, and environmental stress is the major limitation to their productivity. Increasing their tolerance to abiotic and biotic stress is an important goal in agriculture and considerable progress has been made in discovering the mechanisms of cell signaling that regulate the response to these stresses in plants. Signal localization, provided by protein-protein interactions, is the basis for much of signaling specificity and protein phosphorylation is the most widely used mechanism. Arabidopsis has played a leading role in this research but the lineages of Arabidopsis and the cereals diverged 240 M years ago, an evolutionary distance almost as long as the 300 M years separating the lineages of chickens and humans. Rice is a model for research in cereals and its importance will soon increase because it will be the first cereal and second plant genome to be sequenced. Concurrently, there have been tremendous advances in methods for discovering protein-protein interactions, particularly in yeast two-hybrid screening and mass spectrometry of proteins. The convergence of genomic sequence, informatics, and protein-protein interaction technologies has created the opportunity to dramatically enhance our understanding of cell signaling in the cereals, using rice as a model system. This project will generate a protein-protein interaction database for leaf and root tissues, with data from a representative member from each sub-family (E<1x10-110) of the rice protein kinases. This information, together with microarray expression profiling and the analysis of selected gene expression knockout mutants, will facilitate the interpretation of phosphorylation-mediated signaling pathways and will be critical in understanding how to improve stress tolerance in the cereals. Other important objectives of the proposal are to enhance the PlantsP database and sponsor the annual plant phosphorylation meeting, which will accelerate the scientific community's progress in deciphering signaling in plants. Finally, we will use yeast two-hybrid screens and the PlantsP database as the foundation of a minority-focused educational outreach program that uniquely allows students to make real scientific discoveries.


| PlantsP | Top | Abstract | Objectives | Deliverables | Targets | Private materials |

Objectives

The specific objectives of this proposal are:
  1. Analyze the rice genome to identify all protein kinases and isolate 400 representative full-length protein kinase cDNAs and use these in yeast two-hybrid systems and as affinity-tagged cDNAs in transgenic rice plants to identify the kinase-associated proteins in leaf and root tissues.
  2. Functionally characterize knockouts of 100 selected protein kinases to evaluate their biological role.
  3. Enhance the community-oriented PlantsP database to support the experimental data and informatics annotation in this project, and as a future basis for modeling and simulation of regulatory processes in plants.
  4. Community outreach via continued support of the annual plant protein phosphorylation meeting and development of an educational outreach program that involves students in yeast two-hybrid screening of proteins that interact with rice kinases.


| PlantsP | Top | Abstract | Objectives | Deliverables | Targets | Private materials |

Deliverables

  1. Full-length cDNA cloning of 275 cDNAs (Zhu lab)
  2. Rice transformation of 350 constructs (275 tagged & 75 RNAi) (Ronald and Song Labs)
  3. MS proteomics in planta of 275 proteins (Fromm Lab)
  4. Yeast 2-hybrid analyses of 175 kinases (Ronald and Song Labs)

Updated $Date: 2007/10/03 14:24:33 $ by $Author: crystal $