Award Abstract #0321450
Prediction and Validation of Phospho-Regulatory Sites in Crop Plant Proteomes

NSF Org:	IOS Division of Integrative Organismal Systems
Initial Amendment Date:	January 30, 2004
Latest Amendment Date:	January 30, 2004
Award Number:	0321450
Award Instrument:	Standard Grant
Program Manager:	Jane Silverthorne IOS Division of Integrative Organismal Systems BIO Directorate for Biological Sciences
Start Date:	February 1, 2004
Expires:	June 30, 2004 (Estimated)
Awarded Amount to Date:	$691501
Investigator(s):	Jeffrey Harper jfharper@unr.edu (Principal Investigator) Michael Gribskov (Co-Principal Investigator)
Sponsor:	The Scripps Research Institute 10550 N TORREY PINES RD LA JOLLA, CA 92037 / -
NSF Program(s):	PLANT GENOME RESEARCH PROJECT
Field Application(s):
Program Reference Code(s):	BIOT, 9109
Program Element Code(s):	1329

ABSTRACT

The genomes of vascular plants encode more than 1000 protein kinases. The long-term goal is to

define all the regulatory phosphorylation sites in crop plant proteomes. Such sites represent potential control points for all aspects of plant growth, including responses to biotic and abiotic stress. The focus is to use bioinformatics to predict a subset of potentially important sites of phospho-regulation in the rice proteome, and then use a peptide-based experimental strategy to start validating those predictions. The four Specific Aims are:

1. Improve bioinformatics tools to identify motifs involved in phospho-signaling in plants.

2. Conduct a clustering analysis of all Ser and Thr containing sub-sequences in plant proteomes.

3. Design peptide array platform to profile kinase activities in vitro.

4. Identify peptides that can disrupt in vivo signaling pathways (more than 50 will be tested).

The uniqueness and strength of the proposed research rests upon the underlying rationale that a

computational comparison of two plant proteomes can be used as a 'noise filter' to identify

important sequence motifs that are fundamental to signaling pathways conserved between monocots and dicots. This filtering provides the critical foundation for implementing 'peptide-disruptors' as a powerful new approach to understanding and manipulating the dynamics of the plant cell phosphorylome. DNA constructs for all genes found to encode a 'peptide disruptor' or function as a reporter for a particular kinase activity will be deposited at the ABRC at Ohio State University, and will be freely available.

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