Papers by Juan Diego Vizcaino
Journal of microbiology (Seoul, Korea), 2006
Four different Trichoderma strains, T. harzianum CECT 2413, T. asperellum T53, T. atroviride T11 ... more Four different Trichoderma strains, T. harzianum CECT 2413, T. asperellum T53, T. atroviride T11 and T. longibrachiatum T52, which represent three of the four sections contained in this genus, were transformed by two different techniques: a protocol based on the isolation of protoplasts and a protocol based on Agrobacterium-mediated transformation. Both methods were set up using hygromycin B or phleomycin resistance as the selection markers. Using these techniques, we obtained phenotypically stable transformants of these four different strains. The highest transformation efficiencies were obtained with the T. longibrachiatum T52 strain: 65-70 transformants/microg DNA when transformed with the plasmid pAN7-1 (hygromycin B resistance) and 280 transformants/107 spores when the Agrobacterium-mediated transformation was performed with the plasmid pUR5750 (hygromycin B resistance). Overall, the genetic analysis of the transformants showed that some of the strains integrated and maintained...
Proteomics, 2013
The Human Proteome Organisation Proteomics Standards Initiative (HUPO-PSI) was established in 200... more The Human Proteome Organisation Proteomics Standards Initiative (HUPO-PSI) was established in 2002 with the aim of defining community standards for data representation in proteomics and facilitating data comparison, exchange and verification. The 2013 annual spring workshop was hosted by the University of Liverpool, UK and concentrated on updating and refining the existing standards in the light of new methodologies and technologies. To control the inflation of file sizes, strategies for file compression, particularly for mzML files, were explored. Best practices for encoding information such as protein grouping and PTM localisation were refined and documented. Additional example files for the mzQuantML format were designed to provide support for selected reaction monitoring techniques. Enhancements to the PSI Common Query Interface (PSICQUIC) and PSI-MI XML were discussed. Finally, the group engaged in discussion on how the existing work of the HUPO-PSI can be leveraged by the Metabolomics Standards Initiative to improve the capture of metabolite data.
Inferring which protein species have been detected in bottom-up proteomics experiments has been a... more Inferring which protein species have been detected in bottom-up proteomics experiments has been a challenging problem for which solutions have been maturing over the past decade. While many inference approaches now function well in isolation, comparing and reconciling the results generated across different tools remains difficult. It presently stands as one of the greatest barriers in collaborative efforts such as the Human Proteome Project and public repositories like the PRoteomics IDEntifications (PRIDE) database. Here we present a framework for reporting protein identifications that seeks to improve capabilities for comparing results generated by different inference tools. This framework standardizes the terminology for describing protein identification results, associated with the HUPO-Proteomics Standards Initiative (PSI) mzIdentML standard, while still allowing for differing methodologies to reach that final state. It is proposed that developers of software for reporting identification results will adopt this terminology in their outputs. While the new terminology does not require any changes to the core mzIdentML model, it represents a significant change in practice, and, as such, the rules will be released via a new version of the mzIdentML specification (version 1.2) so that consumers of files are able to determine whether the new guidelines have been adopted by export software.
Encyclopedia of Systems Biology, 2013
Methods in molecular biology (Clifton, N.J.), 2013
Both the existence of data standards and public databases or repositories have been key factors b... more Both the existence of data standards and public databases or repositories have been key factors behind the development of the existing "omics" approaches. In this book chapter we first review the main existing mass spectrometry (MS)-based proteomics resources: PRIDE, PeptideAtlas, GPMDB, and Tranche. Second, we report on the current status of the different proteomics data standards developed by the Proteomics Standards Initiative (PSI): the formats mzML, mzIdentML, mzQuantML, TraML, and PSI-MI XML are then reviewed. Finally, we present an easy way to query and access MS proteomics data in the PRIDE database, as a representative of the existing repositories, using the workflow management system (WMS) tool Taverna. Two different publicly available workflows are explained and described.
Proteomics, Jan 5, 2014
Recent advances in high-throughput experimental techniques have led to an exponential increase in... more Recent advances in high-throughput experimental techniques have led to an exponential increase in both the size and the complexity of the datasets commonly studied in biology. Data visualisation is increasingly used as the key to unlock this data, going from hypothesis generation to model evaluation and tool implementation. It is becoming more and more the heart of bioinformatics workflows, enabling scientists to reason and communicate more effectively. In parallel, there has been a corresponding trend towards the development of related software, which has triggered the maturation of different visualisation libraries and frameworks. For bioinformaticians, scientific programmers and software developers, the main challenge is to pick out the most fitting one(s) to create clear, meaningful and integrated data visualisation for their particular use cases. In this review, we introduce a collection of open source or free to use libraries and frameworks for creating data visualisation, cov...
Methods in molecular biology (Clifton, N.J.), 2011
This chapter describes how a pipeline for the analysis of expressed sequence tag (EST) data can b... more This chapter describes how a pipeline for the analysis of expressed sequence tag (EST) data can be -implemented, based on our previous experience generating ESTs from Trichoderma spp. We focus on key steps in the workflow, such as the processing of raw data from the sequencers, the clustering of ESTs, and the functional annotation of the sequences using BLAST, InterProScan, and BLAST2GO. Some of the steps require the use of intensive computing power. Since these resources are not available for small research groups or institutes without bioinformatics support, an alternative will be described: the use of distributed computing resources (local grids and Amazon EC2).
PROTEOMICS, 2008
Large-scale and high-throughput proteomics experiments of specific samples provide substantial am... more Large-scale and high-throughput proteomics experiments of specific samples provide substantial amounts of identified proteins and peptides, which increasingly find their way into centralized, public data repositories. These data typically have potential beyond the analyses performed by the original authors, and can therefore provide considerable added value by being reused for specific, unexplored enquiries. We here reanalyze two CNS-related proteomics datasets, one from the HUPO's Brain Proteome Project, and one from a comprehensive analysis of cerebrospinal fluid in light of the expression of specific splice isoforms from CNS-related genes. We also evaluate the empirically observed peptides of interest against predictions of their proteotypic character.
PROTEOMICS, 2012
The Human Proteome Organisation Proteomics Standards Initiative (HUPO-PSI) was established in 200... more The Human Proteome Organisation Proteomics Standards Initiative (HUPO-PSI) was established in 2002 with the aim of defining community standards for data representation in proteomics and facilitating data comparison, exchange and verification. Over the last 10 years significant advances have been made, with common data standards now published and implemented in the field of both mass spectrometry and molecular interactions. The 2012 meeting further advanced this work, with the mass spectrometry groups finalising approaches to capturing the output from recent developments in the field, such as quantitative proteomics and SRM. The molecular interaction group focused on improving the integration of data from multiple resources. Both groups united with a guest work track, organized by the HUPO Technology/Standards Committee, to formulate proposals for data submissions from the HUPO Human Proteome Project and to start an initiative to collect standard experimental protocols.
Nucleic Acids Research, 2012
The Protein Identifier Cross-Reference (PICR) service is a tool that allows users to map protein ... more The Protein Identifier Cross-Reference (PICR) service is a tool that allows users to map protein identifiers, protein sequences and gene identifiers across over 100 different source databases. PICR takes input through an interactive website as well as Representational State Transfer (REST) and Simple Object Access Protocol (SOAP) services. It returns the results as HTML pages, XLS and CSV files. It has been in production since 2007 and has been recently enhanced to add new functionality and increase the number of databases it covers. Protein subsequences can be Basic Local Alignment Search Tool (BLAST) against the UniProt Knowledgebase (UniProtKB) to provide an entry point to the standard PICR mapping algorithm. In addition, gene identifiers from UniProtKB and Ensembl can now be submitted as input or mapped to as output from PICR. We have also implemented a 'best-guess' mapping algorithm for UniProt. In this article, we describe the usefulness of PICR, how these changes have been implemented, and the corresponding additions to the web services. Finally, we explain that the number of source databases covered by PICR has increased from the initial 73 to the current 102. New resources include several new species-specific Ensembl databases as well as the Ensembl Genome ones.
Nature Biotechnology, 2014
Molecular & Cellular Proteomics, 2011
In proteomics, protein identifications are reported and stored using an unstable reference system... more In proteomics, protein identifications are reported and stored using an unstable reference system: protein identifiers. These proprietary identifiers are created individually by every protein database and can change or may even be deleted over time. To estimate the effect of the searched protein sequence database on the long-term storage of proteomics data we analyzed the changes of reported protein identifiers from all public experiments in the Proteomics Identifications (PRIDE) database by November 2010. To map the submitted protein identifier to a currently active entry, two distinct approaches were used. The first approach used the Protein Identifier Cross Referencing (PICR) service at the EBI, which maps protein identifiers based on 100% sequence identity. The second one (called logical mapping algorithm) accessed the source databases and retrieved the current status of the reported identifier. Our analysis showed the differences between the main protein databases (International Protein Index (IPI), Uni-Prot Knowledgebase (UniProtKB), National Center for Biotechnological Information nr database (NCBI nr), and Ensembl) in respect to identifier stability. For example, whereas 20% of submitted IPI entries were deleted after two years, virtually all UniProtKB entries remained either active or replaced. Furthermore, the two mapping algorithms produced markedly different results. For example, the PICR service reported 10% more IPI entries deleted compared with the logical mapping algorithm. We found several cases where experiments contained more than 10% deleted identifiers already at the time of publication. We also assessed the proportion of peptide identifications in these data sets that still fitted the originally identified protein sequences. Finally, we performed the same overall analysis on all records from IPI, Ensembl, and UniProtKB: two releases per year were used, from 2005. This analysis showed for the first time the true effect of changing protein identifiers on proteomics data. Based on these findings, UniProtKB seems the best database for applications that rely on the long-term storage of proteomics data. Molecular &
Journal of Chromatography A, 2000
The colour pigments of Trichoderma harzianum fermentation broth were separated and the main fract... more The colour pigments of Trichoderma harzianum fermentation broth were separated and the main fractions were tentatively identified by reversed-phase thin-layer chromatography–Fourier transform infrared spectroscopy (RP-TLC–FT-IR), RP-HPLC–diode array detection and RP-HPLC–MS. It was established that the multistep gradient elution developed for RP-TLC separation of pigments can be successfully used as a pilot method for the rational design of gradient elution in
Molecular & cellular proteomics : MCP, 2012
We report the release of mzIdentML, an exchange standard for peptide and protein identification d... more We report the release of mzIdentML, an exchange standard for peptide and protein identification data, designed by the Proteomics Standards Initiative (PSI). The format was developed by the PSI in collaboration with instrument and software vendors, and the developers of the major open-source projects in proteomics. Software implementations have been developed to enable conversion from most popular proprietary and open-source formats, and mzIdentML will soon be supported by the major public repositories. These developments enable proteomics scientists to start working with the standard for exchanging and publishing data sets in support of publications and they provide a stable platform for bioinformatics groups and commercial software vendors to work with a single file format for identification data.
Fungal Genetics and Biology, 2007
In the present article, we describe the cloning and characterization of the Trichoderma harzianum... more In the present article, we describe the cloning and characterization of the Trichoderma harzianum hmgR gene encoding a hydroxymethylglutaryl CoA reductase (HMGR), a key enzyme in the biosynthesis of terpene compounds. In T. harzianum, partial silencing of the hmgR gene gave rise to transformants with a higher level of sensitivity to lovastatin, a competitive inhibitor of the HMGR enzyme. In addition, these hmgR-silenced transformants produced lower levels of ergosterol than the wild-type strain in a minimal medium containing lovastatin. The silenced transformants showed a decrease in hmgR gene expression (up to a 8.4-fold, after 72 h of incubation), together with an increase in the expression of erg7 (up to a 15.8-fold, after 72 h of incubation), a gene involved in the biosynthesis of triterpenes. Finally, hmgR-silenced transformants showed a reduction in their antifungal activity against the plant-pathogen fungi Rhizoctonia solani and Fusarium oxysporum.
Fungal Genetics and Biology, 2006
Trichoderma species are commonly used as biocontrol agents of diVerent plant-pathogenic fungi. Te... more Trichoderma species are commonly used as biocontrol agents of diVerent plant-pathogenic fungi. Terpene compounds are involved in the biocontrol process due to their antifungal properties (e.g., ergokonins and viridins) but additionally their structural function in the cell membranes (ergosterol) is essential. We report here the characterization of the T. harzianum erg1 gene, encoding a squalene epoxidase, a key enzyme in the biosynthesis of triterpene derivatives such as ergosterol. In T. harzianum the partial silencing of the erg1 gene gave rise to transformants with a higher level of sensitivity to terbinaWne, an antifungal compound that acts speciWcally over the squalene epoxidase activity. In addition, these silenced transformants produced lower levels of ergosterol than the wild type strain. Finally, the silencing of the erg1 gene resulted in an increase in the expression level of the erg7 gene that encodes the oxidosqualene lanosterol-cyclase, another enzyme of the terpene biosynthesis pathway.
Mycological Research, 2004
Genetic variability within 69 biocontrol isolates of Trichoderma, obtained from different geograp... more Genetic variability within 69 biocontrol isolates of Trichoderma, obtained from different geographical locations and culture collections and selected as biocontrol agents, was studied. Sequence data, obtained from the ITS1 region of rDNA and a fragment of the translation elongation factor 1 (tef1) gene, were used in a phylogenetic analysis. Phylograms showing similar topologies were generated using alignments containing the ITS1 region or a portion of the tef1 gene. 21 distinct ITS1 sequence types and 17 distinct tef1 sequence types were identified among the 69 isolates. More than 50 % of the potential biocontrol strains were grouped within Trichoderma sect. Pachybasium ; of these, 81 % were grouped within the cluster that included the ex-type strains of T. harzianum and T. inhamatum, and 16 % were grouped with T. virens. Within T. sect. Trichoderma, which included 36 % of the 69 strains, 56 % were grouped with T. asperellum, and 24 % with T. viride, T. atroviride or T. koningii. Only 10 % of the strains studied were located in T. sect. Longibrachiatum.
Mycological Research, 2005
Methanol extracts from 24 Trichoderma isolates, selected as biocontrol agents and representating ... more Methanol extracts from 24 Trichoderma isolates, selected as biocontrol agents and representating different species and genotypes from three of the four taxonomic sections of this genus (T. sect. Trichoderma, T. sect. Pachybasium and T. sect. Longibrachiatum) were screened for antibacterial, anti-yeast and antifungal activities against a panel of seven bacteria, seven yeasts and six filamentous fungi previously used in similar studies. Two different growth media were tested (potato dextrose broth and CYS80), and all isolates included in the antimicrobial tests showed at least one inhibitory activity against one of the target microorganisms in one of the two culture media. No statistically significant differences were detected in the number of active strains between the two culture media, but the highest number of inhibitory strains against bacteria and fungi were found in strains from Trichoderma sect. Pachybasium, whereas strains from T. sect. Longibrachiatum showed the highest anti-yeast values. In all cases, a correlation was found between the strains that were active against yeasts and fungi. However, some degree of variability was detected for strains within the same taxonomic section. In general terms, strains from T. asperellum (mainly in CYS80 medium), and T. longibrachiatum gave the best non-enzymatic antimicrobial profiles.
FEMS Microbiology Letters, 2005
Some of the secondary metabolites produced by Trichoderma, such as the peptaibols and other antib... more Some of the secondary metabolites produced by Trichoderma, such as the peptaibols and other antibiotics, have a peptide structure and in their biosynthesis are involved proteins belonging to the Non-Ribosomal Peptide Synthetase family. In the present work, a PCR-mediated strategy was used to clone a region corresponding to an adenylation domain of a peptide synthetase (PS) gene from 10 different strains of Trichoderma. In addition, and using the fragment isolated by PCR from T. harzianum CECT 2413 as a probe, a fragment of 19.0 kb corresponding to a PS-encoding gene named salps1, including a 1.5 kb fragment of the promoter, was cloned and sequenced. The cloned region of salps1 contains four complete, and a fifth incomplete, modules, in which are found the adenylation, thiolation and condensation domains, but also an additional epimerization domain at the C-terminal end of the first module. The analysis of the Salps1 protein sequence, taking into consideration published data, suggests that it is neither a peptaibol synthetase nor a protein involved in siderophore biosynthesis. The presence of two breaks in the open reading frame and the expression of this gene under nitrogen starvation conditions suggest that salps1 could be a pseudogene.
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Papers by Juan Diego Vizcaino