Theses and Dissertations

Issuing Body

Mississippi State University

Advisor

Li, Jiaxu

Committee Member

Peterson, Daniel G.

Committee Member

Ma, Din-Pow

Committee Member

Willard, Scott

Committee Member

Peng, Zhaohua

Date of Degree

8-14-2015

Document Type

Dissertation - Open Access

Major

Molecular Biology

Degree Name

Doctor of Philosophy

College

College of Agriculture and Life Sciences

Department

Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology

Abstract

Plant hormone abscisic acid (ABA) plays a main role in coordinating various stress signals in plants. ABA regulates the expression of genes and activities of enzymes in response to various stress conditions. In the following studies we were able to study the ABA mediated regulation of enzymes in plants. Using in-gel activity analysis we identified that ABA regulates the activity of aspartate aminotransferase (AAT), an enzyme involved in nitrogen assimilation and carbohydrate metabolism. Our results indicate that phosphorylation of AAT by SnRK2.2 and 2.3 kinases results in down regulation of AAT2 and AAT3 isozyme activities in Arabidopsis. AAT was identified as a negative regulator of drought stress and aat mutant plants showed improved survival following drought conditions. Using in-gel staining method we were able to visualize sugar phosphatases like fructose 1-6 bisphosphatase family, sedoheptulase-1,7-bisphosphatase, inositol mono phosphatases; protein serine/threonine phosphatases, protein tyrosine phosphatases and studied their response to ABA and drought stress. Fructose-1-6 bisphosphatase family of phosphatases were identified to be induced by ABA in Arabidopsis and rice. N-acetylglucosamine (GlcNAc) is present on glycoproteins and as post translational modification (PTM) in cytoplasmic and nuclear proteins. N-acetylglucosamine is removed from target proteins by hexosaminidases. Little is known about the hexosaminidases in plants. Using in-gel activity analysis we were able to identify an ABA induced Beta-hexosaminidase with a neutral pH optimum in soybean. The nuclear DNA in chromatin is associated with basic proteins called histones. The N-terminal tails of histones contain different PTMs including methylation, phosphorylation, ubiquitination, acetylation, ADP-ribosylation and glycosylation. The histone lysine methylation can serve as a binding site or repel/disrupt the histone binding proteins. The effector/reader proteins specifically recognize the post translational modifications and responsible for the downstream process. Many histone methyl modification effector proteins have been characterized but very few proteins whose binding was disrupted by the presence of a PTM were identified. Using peptide pulldown analysis, far western analyses we identified a WD-40 domain containing histone binding (HB01) protein as direct interactor of unmodified histone. The presence of post translational modifications disrupts HB01 binding to histone H3.

URI

https://hdl.handle.net/11668/17025

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