Award Abstract #0115937
Arabidopsis 2010: Collaborative Project on the Functional Genomics of Arabidopsis beta-Glucosidase and beta-Galactosidase Gene Families

NSF Org:	MCB Division of Molecular and Cellular Biosciences
Initial Amendment Date:	September 21, 2001
Latest Amendment Date:	June 27, 2006
Award Number:	0115937
Award Instrument:	Continuing grant
Program Manager:	David A. Rockcliffe MCB Division of Molecular and Cellular Biosciences BIO Directorate for Biological Sciences
Start Date:	September 15, 2001
Expires:	August 31, 2007 (Estimated)
Awarded Amount to Date:	$1135250
Investigator(s):	Asim Esen aevatan@vt.edu (Principal Investigator) Brenda Winkel (Co-Principal Investigator) David Bevan (Co-Principal Investigator)
Sponsor:	Virginia Polytechnic Institute and State University 1880 Pratt Drive BLACKSBURG, VA 24060 540/231-5281
NSF Program(s):	MOLECULAR BIOCHEMISTRY, BIOMOLECULAR SYSTEMS
Field Application(s):
Program Reference Code(s):	SMET, BIOT, 9251, 9179, 9178, 9109, 1684, 1228, 1166
Program Element Code(s):	1166, 1144

ABSTRACT

Widely distributed in animals, plants and microbes, O-glycoside hydrolases (EC 3.2.1.-) are enzymes that catalyze the cleavage of chemical bonds between two or more carbohydrates or between a carbohydrate and a non-carbohydrate moiety. Responding to the challenge of the 2010 Project to identify the function of all Arabidopsis thaliana genes within the next decade, this collaborative research will focus on approximately 75 members of two related families of glycoside hydrolases (http://www.biol.vt.edu/faculty/esen/glycosidase.lab.html and http://afmb.cnrs-mrs.fr/~pedro/CAZY/). Family 1 includes beta -glucosidases (EC 3.2.1.21) and myrosinases (EC 3.2.3.1), which function in higher plants in chemical defense against herbivores and pathogens, lignin biosynthesis, and plant growth and development. Family 35 contains the beta -galactosidases (EC 3.2.1.23), which play key roles in fruit ripening, flower senescence, mobilization of carbohydrate reserves, and galactolipid turnover. To date, the precise biochemical roles of only three of these Arabidopsis hydrolases are known with certainty.

The purpose of this multidisciplinary collaborative research is to assign biological functions to beta-glucosidases and beta-galactosidases encoded by the Arabidopsis genome. After using phylogenetic analysis to identify subfamilies that contain closely related enzymes, cDNAs encoding each target hydrolase will be obtained. Each hydrolase will then be overexpressed in foreign host (e.g. yeast and bacteria) cells and purified to ascertain its biological function by measuring its enzymatic activity toward a wide range of natural glycosidic substrates isolated primarily from Arabidopsis and related crucifers for this purpose. In parallel studies, three-dimensional structures of selected subfamily representatives will be determined by homology modeling and x-ray diffraction, providing novel insights into how these hydrolases recognize and bind their substrates. This information will be of paramount importance in future research to alter the substrate specificity of Family 1 and Family 35 hydrolases for biotechnological purposes, including biomass conversion and improvements in anti-herbivore defenses and fruit ripening.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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Ahn, Y. O; Zheng, M; Bevan, D. R; Esen, A; Shiu, S-H; Benson, J; Peng, H-P; Miller, J. T;. Cheng, C-L; Poulton, J.E; Shih, M-C.  "Functional genomic analysis of Arabidopsis thaliana glycoside hydrolase family 35,"  Phytochemistry,  v.68,  2007,  p. 1510.

Esen, A., Jonathan E. Poulton, C.-L. Cheng, M.-C. Shih, A. Mohamed, B. Winkel-Shirley, D. R. Bevan, B. Henrissat, B. L. M°ller.  "Collaborative Project on the Functional Genomics of the Arabidopsis beta-Glucosidase and beta-Galactosidase Gene Families. In: F. M. Ausubel ôSummaries of National Science Foundation-Sponsored Arabidopsis 2010 Projects,"  Plant Physiology,  v.129,  2002,  p. 394.

Xu, Z.; Escamilla-Trevino, L; Zeng, L; Lalgondar, M; Bevan, D. R; B. S. Winkel, B. S; Mohamed, A; Cheng, C. L; Shih, M.-C; Poulton, J.E; Esen, A.  "Functional genomic analysis of Arabidopsis thaliana glycoside hydrolase family 1,"  Plant Molecular Biology,  v.55,  2004,  p. 343.


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