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

Autor:	Chen Y; GBF, Université de Toulouse, INRA, Toulouse, France., Rofidal V; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Hem S; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Gil J; GBF, Université de Toulouse, INRA, Toulouse, France.; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Nosarzewska J; GBF, Université de Toulouse, INRA, Toulouse, France., Berger N; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Demolombe V; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Bouzayen M; GBF, Université de Toulouse, INRA, Toulouse, France., Azhar BJ; Department of Biochemistry, Quaid-i-azam University, Islamabad, Pakistan.; Department of Biological Sciences, Dartmouth College, Hanover, NH, United States., Shakeel SN; Department of Biochemistry, Quaid-i-azam University, Islamabad, Pakistan.; Department of Biological Sciences, Dartmouth College, Hanover, NH, United States., Schaller GE; Department of Biological Sciences, Dartmouth College, Hanover, NH, United States., Binder BM; Department of Biochemistry, Cellular, and Molecular Biology, University of Tennessee, Knoxville, TN, United States., Santoni V; BPMP, CNRS, INRA, Montpellier SupAgro, Université de Montpellier, Montpellier, France., Chervin C; GBF, Université de Toulouse, INRA, Toulouse, France.
Jazyk:	angličtina
Zdroj:	Frontiers in plant science [Front Plant Sci] 2019 Aug 29; Vol. 10, pp. 1054. Date of Electronic Publication: 2019 Aug 29 (Print Publication: 2019).
DOI:	10.3389/fpls.2019.01054
Abstrakt:	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.
Databáze:	MEDLINE
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