Dataset Information

Saprolegnia parasitica proteome

ABSTRACT: In this project, we made a comprehensive quantitative proteomic analysis of four defined developmental stages of S. parasitica (mycelium, primary cysts, secondary cysts and germinated cysts) to gain greater insight into the types of proteins specifically linked to each stage. A total of 2423 unique proteins were identified using qualitative (gel-based) and quantitative (iTRAQ, isobaric tags for relative and absolute quantitation) approaches. Of these, 358 proteins, associated with various biological processes, were found to be significantly enriched between different life cycle stages of S. parasitica. The transcript abundance of several cyst and mycelium enriched proteins was also checked by quantitative real-time PCR. This is the first large scale proteomic analysis on Saprolegnia and the data from this study will enhance our current knowledge about this pathogen by identifying biological processes key to each developmental stage.

INSTRUMENT(S): Xevo Q-Tof

ORGANISM(S): Oomycetes

TISSUE(S): Cyst

SUBMITTER: Vaibhav Srivastava

LAB HEAD: Vincent Bulone / Vaibhav Srivastava

PROVIDER: PXD004695 | Pride | 2018-08-10

REPOSITORIES: Pride

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Publications

Quantitative Proteomic Analysis of Four Developmental Stages of Saprolegnia parasitica.

Srivastava Vaibhav V Rezinciuc Svetlana S Bulone Vincent V

Frontiers in microbiology 20180111

Several water mold species from the Saprolegnia genus infect fish, amphibians, and crustaceans in natural ecosystems and aquaculture farms. Saprolegnia parasitica is one of the most severe fish pathogens. It is responsible for millions of dollars of losses to the aquaculture industry worldwide. Here, we have performed a proteomic analysis, using gel-based and solution (iTRAQ) approaches, of four defined developmental stages of S. parasitica grown in vitro, i.e., the m ...[more]

PMID: 29375523

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Project description:The recent development within high-throughput technologies for expression profiling has allowed for parallel analysis of transcriptomes and proteomes in biological systems such as comparative analysis of transcript and protein levels of tissue regulated genes. Until now, such studies of have only included microarray or short length sequence tags for transcript profiling. Furthermore, most comparisons of transcript and protein levels have been based on absolute expression values from within the same tissue and not relative expression values based on tissue ratios. Presented here is a novel study of two porcine tissues based on integrative analysis of data from expression profiling of identical samples using cDNA microarray, 454-sequencing and iTRAQ-based proteomics. Sequence homology identified 2.541 unique transcripts that are detectable by both microarray hybridizations and 454-sequencing of 1.2 million cDNA tags. Both transcript-based technologies showed high reproducibility between sample replicates of the same tissue, but the correlation across these two technologies was modest. Thousands of genes being differentially expressed were identified with microarray. Out of the 306 differentially expressed genes, identified by 454-sequencing, 198 (65 %) were also found by microarray. The relationship between the regulation of transcript and protein levels was analyzed by integrating iTRAQ-based proteomics data. Protein expression ratios were determined for 354 genes, of which 148 could be mapped to both microarray and 454-sequencing data. A comparison of the expression ratios from the three technologies revealed that differences in transcript and protein levels across heart and muscle tissues are positively correlated. We show that the reproducibility within cDNA microarray and 454-sequencing is high, but that the agreement across these two technologies is modest. We demonstrate that the regulation of transcript and protein levels across identical tissue samples is positively correlated when the tissue expression ratios are used for comparison. The results presented are of interest in systems biology research in terms of integration and analysis of high-throughput expression data from mammalian tissues. Keywords: tissue comparison, platform comparison Tissue samples of heart (HEA) and Longissimus dorsi (LDO) were prepared by pooling from five healthy Hampshire gilts at age four to six months and division into six sub-samples, three for each tissue named HEA1, HEA2, HEA3, LDO1, LDO2 and LDO3. The exact same six tissue samples were used for subsequent expression profiling with cDNA microarray, 454-sequencing and iTRAQ-based proteomics. A reference sample for the cDNA microarray experiment and iTRAQ-based proteomics was constructed by combining the six samples. In the microarray experiment, three cDNA microarray slides were used per sample.

Dataset Information

Saprolegnia parasitica proteome

Publications

Quantitative Proteomic Analysis of Four Developmental Stages of <i>Saprolegnia parasitica</i>.

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