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Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes

Chen, Yi and Rofidal, Valérie and Hem, Sonia and Gil, Julie and Nosarzewska, Joanna and Berger, Nathalie and Demolombe, Vincent and Bouzayen, Mondher and Azhar, Beenish and Shakeel, Samina and Schaller, Georges Eric and Binder, Brad Mark and Santoni, Véronique and Chervin, Christian Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes. (2019) Frontiers in Plant Science, 10. 1-10. ISSN 1664-462X

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Official URL: https://www.frontiersin.org/articles/10.3389/fpls.2019.01054/full

Abstract

Ethylene regulates fruit ripening and several plant functions (germination, plant growth, plant-microbe interactions). Protein quantification of ethylene receptors (ETRs) is essential to study their functions, but is impaired by low resolution tools such as antibodies that are mostly nonspecific, or the lack of sensitivity of shotgun proteomic approaches. We developed a targeted proteomic method, to quantify low-abundance proteins such as ETRs, and coupled this to mRNAs analyses, in two tomato lines: Wild Type (WT) and Never-Ripe (NR) which is insensitive to ethylene because of a gain-of-function mutation in ETR3. We obtained mRNA and protein abundance profiles for each ETR over the fruit development period. Despite a limiting number of replicates, we propose Pearson correlations between mRNA and protein profiles as interesting indicators to discriminate the two genotypes: such correlations are mostly positive in the WT and are affected by the NR mutation. The influence of putative post-transcriptional and post-translational changes are discussed. In NR fruits, the observed accumulation of the mutated ETR3 protein between ripening stages (Mature Green and Breaker + 8 days) may be a cause of NR tomatoes to stay orange. The label-free quantitative proteomics analysis of membrane proteins, concomitant to Parallel Reaction Monitoring analysis, may be a resource to study changes over tomato fruit development. These results could lead to studies about ETR subfunctions and interconnections over fruit development. Variations of RNA-protein correlations may open new fields of research in ETR regulation. Finally, similar approaches may be developed to study ETRs in whole plant development and plant-microorganism interactions.

Item Type:Article
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Université de Toulouse > Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE)
French research institutions > Institut National de la Recherche Agronomique - INRA (FRANCE)
Other partners > Montpellier SupAgro (FRANCE)
Other partners > Dartmouth College (USA)
Other partners > Quaid-i-Azam University (Pakistan)
Other partners > Université de Montpellier (FRANCE)
Other partners > University of Tennessee - UT (USA)
Laboratory name:
Funders:
National Science Foundation - NSF (USA)
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Deposited On:18 Dec 2019 14:12

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