Publications by authors named "Eva Liñeiro"

7 Publications

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Epistasis-driven identification of SLC25A51 as a regulator of human mitochondrial NAD import.

Nat Commun 2020 12 1;11(1):6145. Epub 2020 Dec 1.

CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.

About a thousand genes in the human genome encode for membrane transporters. Among these, several solute carrier proteins (SLCs), representing the largest group of transporters, are still orphan and lack functional characterization. We reasoned that assessing genetic interactions among SLCs may be an efficient way to obtain functional information allowing their deorphanization. Here we describe a network of strong genetic interactions indicating a contribution to mitochondrial respiration and redox metabolism for SLC25A51/MCART1, an uncharacterized member of the SLC25 family of transporters. Through a combination of metabolomics, genomics and genetics approaches, we demonstrate a role for SLC25A51 as enabler of mitochondrial import of NAD, showcasing the potential of genetic interaction-driven functional gene deorphanization.
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http://dx.doi.org/10.1038/s41467-020-19871-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7708531PMC
December 2020

Proteomic study of the membrane components of signalling cascades of Botrytis cinerea controlled by phosphorylation.

Sci Rep 2019 07 8;9(1):9860. Epub 2019 Jul 8.

Andalusian Center for Grape and Grapevine Research (IVAGRO), Microbiology Lab, University of Cadiz, Puerto Real, 11510, Spain.

Protein phosphorylation and membrane proteins play an important role in the infection of plants by phytopathogenic fungi, given their involvement in signal transduction cascades. Botrytis cinerea is a well-studied necrotrophic fungus taken as a model organism in fungal plant pathology, given its broad host range and adverse economic impact. To elucidate relevant events during infection, several proteomics analyses have been performed in B. cinerea, but they cover only 10% of the total proteins predicted in the genome database of this fungus. To increase coverage, we analysed by LC-MS/MS the first-reported overlapped proteome in phytopathogenic fungi, the "phosphomembranome" of B. cinerea, combining the two most important signal transduction subproteomes. Of the 1112 membrane-associated phosphoproteins identified, 64 and 243 were classified as exclusively identified or overexpressed under glucose and deproteinized tomato cell wall conditions, respectively. Seven proteins were found under both conditions, but these presented a specific phosphorylation pattern, so they were considered as exclusively identified or overexpressed proteins. From bioinformatics analysis, those differences in the membrane-associated phosphoproteins composition were associated with various processes, including pyruvate metabolism, unfolded protein response, oxidative stress response, autophagy and cell death. Our results suggest these proteins play a significant role in the B. cinerea pathogenic cycle.
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http://dx.doi.org/10.1038/s41598-019-46270-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6614480PMC
July 2019

Phenotypic Effects and Inhibition of Botrydial Biosynthesis Induced by Different Plant-Based Elicitors in Botrytis cinerea.

Curr Microbiol 2018 Apr 17;75(4):431-440. Epub 2017 Nov 17.

Microbiology Laboratory, Andalusian Center for Grape and Grapevine Research, Faculty of Marine and Environmental Sciences, University of Cadiz, Polígono Rio San Pedro s/n, 11510, Puerto Real, Cádiz, Spain.

Botrytis cinerea is considered a model organism for the study of plant-pathogen interaction showing great genetic diversity and a high degree of morphological variability depending on environmental conditions. The use of new compounds and plant-based elicitors may trigger the expression of different B. cinerea genes, providing new sources of virulence factors. This work is focused on elucidating the phenotypic effect in B. cinerea of different carbon sources such as glucose, cellulose and tomato cell walls (TCW). Production of botrydial and dihydrobotrydial toxins was evaluated using thin-layer chromatography (TLC), proton nuclear magnetic resonance spectroscopy (H-NMR) and mass spectrometry (UPLC-HRESIMS). Expression of the toxin biosynthesis gene BcBOT2 was followed using RT-qPCR. Results show an inhibition of the toxin biosynthesis pathway when TCW are present as a sole carbon source, suggesting that the toxin is only produced when rich molecules, like glucose, are available for fungal metabolism. That suggests a connection between gene expression of virulence factors and environmental conditions, where the silent genes can be induced by different culture conditions.
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http://dx.doi.org/10.1007/s00284-017-1399-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5842495PMC
April 2018

Dataset of the phosphoproteome induced by different plant-based elicitors.

Data Brief 2016 Jun 22;7:1447-1450. Epub 2016 Apr 22.

Andalusian Center for Grape and Grapevine Research, CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, 11510 Cádiz, Spain.

Phosphorylation is one of the main post-translational modification (PTM) involved in signaling network in the ascomycete , one of the most relevant phytopathogenic fungus. The data presented in this article provided a differential mass spectrometry-based analysis of the phosphoproteome of under two different phenotypical conditions induced by the use of two different elicitors: glucose and deproteinized Tomate Cell Walls (TCW). A total 1138 and 733 phosphoproteins were identified for glucose and TCW culture conditions respectively. Raw data are deposited at the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier (PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD003099). Further interpretation and discussion of these data are provided in our research article entitled "Phosphoproteome analysis of in response to different plant-based elicitors" (Liñeiro et al., 2016) [1].
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http://dx.doi.org/10.1016/j.dib.2016.04.039DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5063813PMC
June 2016

Modifications of fungal membrane proteins profile under pathogenicity induction: A proteomic analysis of Botrytis cinerea membranome.

Proteomics 2016 09 5;16(17):2363-76. Epub 2016 Aug 5.

Andalusian Center for Grape and Grapevine Research (IVAGRO), CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, Spain.

Botrytis cinerea is a model fungus for the study of phytopathogenicity that exhibits a wide arsenal of tools to infect plant tissues. Most of these factors are related to signal transduction cascades, in which membrane proteins play a key role as a bridge between environment and intracellular molecular processes. This work describes the first description of the membranome of Botrytis under different pathogenicity conditions induced by different plant-based elicitors: glucose and tomato cell wall (TCW). A discovery proteomics analysis of membrane proteins was carried out by mass spectrometry. A total of 2794 proteins were successfully identified, 46% of them were classified as membrane proteins based on the presence of transmembrane regions and lipidation. Further analyses showed significant differences in the membranome composition depending on the available carbon source: 804 proteins were exclusively identified when the fungus was cultured with glucose as a sole carbon source, and 251 proteins were exclusively identified with TCW. Besides, among the 1737 common proteins, a subset of 898 proteins presented clear differences in their abundance. GO enrichment and clustering interaction analysis revealed changes in the composition of membranome with increase of signalling function in glucose conditions and carbohydrate degradation process in TCW conditions. All MS data have been deposited in the ProteomeXchange with identifier PXD003099 (http://proteomecentral.proteomexchange.org/dataset/PXD003099).
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http://dx.doi.org/10.1002/pmic.201500496DOI Listing
September 2016

Phosphoproteome analysis of B. cinerea in response to different plant-based elicitors.

J Proteomics 2016 Apr 18;139:84-94. Epub 2016 Mar 18.

Andalusian Center for Grape and Grapevine Research, CeiA3, Marine and Environmental Sciences Faculty, University of Cadiz, Puerto Real, 11510 Cádiz, Spain. Electronic address:

Unlabelled: The phytopathogen Botrytis cinerea is a ubiquitous fungus with a high capacity to adapt its metabolism to different hosts and environmental conditions in order to deploy a variety of virulence and pathogenicity factors and develop a successful plant infection. Here we report the first comparative phosphoproteomic study of B. cinerea, aimed to analyze the phosphoprotein composition of the fungus and its changes under different phenotypical conditions induced by two different carbon sources as plant based elicitors: glucose and deproteinized tomato cell wall (TCW). A total of 2854 and 2269 different phosphosites (2883 and 1137 phosphopeptides) were identified in glucose and TCW respectively, which map to 1338 phosphoproteins in glucose and 733 in TCW. Out of the identified phosphoproteins, 173 were exclusively found when glucose was the only carbon source and 11 when the carbon source was TCW. Differences in the pattern of phosphorylation-sites were also detected according to the carbon source. Gene ontology classification of the identified phosphoproteins showed that most of the characteristic proteins of the different carbon sources were related to signalling and transmembrane transport, thus highlighting the importance of these processes in the fungal adaptation to the surrounding conditions.

Biological Significance: The characterization of the B. cinerea phosphoproteome under different induction conditions reported here is the first comparative phosphoproteomic approach in this model phytopathogenic fungus. The identified phosphopeptides contribute to expand the map of known phosphoproteins in this pathogen and the observed changes according to the used carbon source contribute to understand the adaptation of the fungus to the environment changes. This knowledge improves the understanding of the adaptation mechanism, defines the role of the phosphoproteins involved in this process, and enables the advance in the design of novel strategies against the fungi.
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http://dx.doi.org/10.1016/j.jprot.2016.03.019DOI Listing
April 2016

Proteomic profiling of Botrytis cinerea conidial germination.

Arch Microbiol 2015 Mar 21;197(2):117-33. Epub 2014 Aug 21.

Laboratory of Microbiology, Marine and Environmental Sciences Faculty, Andalusian Center for Grape and Grapevine Research, CeiA3 International Campus of Excellence in Agrifood, University of Cádiz, Pol. Río San Pedro s/n, 11510, Puerto Real, Cádiz, Spain.

Botrytis cinerea is one of the most relevant plant pathogenic fungi. The first step during its infection process is the germination of the conidia. Here, we report on the first proteome analysis during the germination of B. cinerea conidia, where 204 spots showed significant differences in their accumulation between ungerminated and germinated conidia by two-dimensional polyacrylamide gel electrophoresis and qPCR. The identified proteins were grouped by gene ontology revealing that the infective tools are mainly preformed inside the ungerminated conidia allowing a quick fungal development at the early stages of conidial germination. From 118 identified spots, several virulence factors have been identified while proteins, such as mannitol-1-phosphate dehydrogenase, 6,7-dimethyl-8-ribityllumazine synthase or uracil phosphoribosyltransferase, have been disclosed as a new potential virulence factors in botrytis whose role in pathogenicity needs to be studied to gain new insights about the role of these proteins as therapeutic targets and virulence factors.
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http://dx.doi.org/10.1007/s00203-014-1029-4DOI Listing
March 2015