Proteomic Analysis of the Function of a Novel Cold-Regulated Multispanning Transmembrane Protein COR413-PM1 in Arabidopsis.

Chen Su,Youzhi Ma,Qingqian Ding,Zhaoshi Xu,Ming Chen,Yongying Mou,Rui Yang,Yinghong Pan,Yajun Xi,Mengjie Zhao,Bo Ma,Kai Chen

doi:10.3390/ijms19092572

Abstract

The plasma membrane is the first subcellular organ that senses low temperature, and it includes some spanning transmembrane proteins that play important roles in cold regulation. COR413-PM1 is a novel multispanning transmembrane cold-regulated protein; however, the related functions are not clear in Arabidopsis. We found the tolerance to freezing stress of cor413-pm1 was lower than wild-type (WT). A proteomics method was used to analyze the differentially abundant proteins (DAPs) between cor413-pm1 and WT. A total of 4143 protein groups were identified and 3139 were accurately quantitated. The DAPs associated with COR413-PM1 and freezing treatment were mainly involved in the metabolism of fatty acids, sugars, and purine. Quantitative real-time PCR (qRT-PCR) confirmed the proteomic analysis results of four proteins: fatty acid biosynthesis 1 (FAB1) is involved in fatty acid metabolism and might affect the plasma membrane structure; fructokinase 3 (FRK3) and sucrose phosphate synthase A1 (SPSA1) play roles in sugar metabolism and may influence the ability of osmotic adjustment under freezing stress; and GLN phosphoribosyl pyrophosphate amidotransferase 2 (ASE2) affects freezing tolerance through purine metabolism pathways. In short, our results demonstrate that the multispanning transmembrane protein COR413-PM1 regulates plant tolerance to freezing stress by affecting the metabolism of fatty acids, sugars, and purine in Arabidopsis.

Highlights

Cold stress tolerance is essential for plant development and environmental acclimation
To identify the homozygous genotype of the transfer DNA (T-DNA) insertion mutant cor413-pm1, the DNA extractions of the mutant cor413-pm1 and WT were used as templates for amplification using the insertion site-specific primers of cor413-pm1 (T-DNA was inserted into the promoter region of COR413-PM1 gene shown in Figure 1B) and T-DNA border primer (LBb1.3) [28,29]
Results from the Quantitative real-time PCR (qRT-PCR) implied that the expression of genes were diminished in the mutant cor413-pm1, which confirmed the results of the proteomic analysis and suggested that proteins might be implicated in increasing plant tolerance to cold

Summary

Introduction

Cold stress tolerance is essential for plant development and environmental acclimation. Significant progress has been made in the past several decades in describing the plant transcriptome regulating network associated with cold stress from exposure to chilling and freezing temperatures, and this work has significantly contributed to our understanding of tolerance mechanisms to cold stress [1]. The CBF transcription factors (CBF1/DREB1b, CBF2/DREB1c, and CBF3/DREB1a) together with an APETALA2/ethylene response factor (AP2) DNA binding domain play a key role in the cold signaling pathway in plants [6,7]. This signal transduction causes a series of downstream responses to cold. Plants have evolved various mechanisms for cold sensing and stress signal transduction, and these varied mechanisms interact to produce cold tolerance

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Discussion

Conclusion