Proteomics Approach to Identify Dehydration Responsive Nuclear Proteins from Chickpea (Cicer arietinum L.)

Aarti Pandey,Subhra Chakraborty,Asis Datta,Niranjan Chakraborty

doi:10.1074/mcp.m700314-mcp200

Aarti Pandey, Subhra Chakraborty + Show 2 more

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https://doi.org/10.1074/mcp.m700314-mcp200

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Abstract

Dehydration or water-deficit is one of the most important environmental stress factors that greatly influences plant growth and development and limits crop productivity. Plants respond and adapt to such stress by altering their cellular metabolism and activating various defense machineries. Mechanisms that operate signal perception, transduction, and downstream regulatory events provide valuable information about the underlying pathways involved in environmental stress responses. The nuclear proteins constitute a highly organized, complex network that plays diverse roles during cellular development and other physiological processes. To gain a better understanding of dehydration response in plants, we have developed a comparative nuclear proteome in a food legume, chickpea (Cicer arietinum L.). Three-week-old chickpea seedlings were subjected to progressive dehydration by withdrawing water and the changes in the nuclear proteome were examined using two-dimensional gel electrophoresis. Approximately 205 protein spots were found to be differentially regulated under dehydration. Mass spectrometry analysis allowed the identification of 147 differentially expressed proteins, presumably involved in a variety of functions including gene transcription and replication, molecular chaperones, cell signaling, and chromatin remodeling. The dehydration responsive nuclear proteome of chickpea revealed a coordinated response, which involves both the regulatory as well as the functional proteins. This study, for the first time, provides an insight into the complex metabolic network operating in the nucleus during dehydration.

Highlights

Dehydration or water-deficit is one of the most important environmental stress factors that greatly influences plant growth and development and limits crop productivity
The nuclear proteins were prepared from the purified nuclei using TriPure reagent (Roche Diagnostics) to remove the contaminating nucleic acids, which might interfere during the isoelectric focusing (IEF) process
The enrichment of nuclear proteins was evaluated by immunoblot analysis using specific antibodies for two nuclear proteins, fibrillarin and histone core (Fig. 1B)

Summary

Introduction

Dehydration or water-deficit is one of the most important environmental stress factors that greatly influences plant growth and development and limits crop productivity. To gain a better understanding of dehydration response in plants, we have developed a comparative nuclear proteome in a food legume, chickpea (Cicer arietinum L.). The dehydration responsive nuclear proteome of chickpea revealed a coordinated response, which involves both the regulatory as well as the functional proteins. Nuclear proteins play key roles in the fundamental regulation of genome instability, the phases of organ development, and physiological responsiveness through gene expression. We have reported the nucleus-specific comparative proteome of chickpea to identify novel components involved in dehydration tolerance with a wider aim to use them in future crop improvement program. The comparison of dehydration responsive nuclear proteome in chickpea reveals predicted and unexpected components indicating their possible role in dehydration tolerance

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