A parallel proteomic and metabolomic analysis of the hydrogen peroxide- and Sty1p-dependent stress response in Schizosaccharomyces pombe

• Published in: Proteomics (Weinheim) Vol. 6; no. 9; pp. 2772 - 2796
• Main Authors: Weeks, Mark E., Sinclair, John, Butt, Amna, Chung, Yuen-Li, Worthington, Jessica L., Wilkinson, Caroline R. M., Griffiths, John, Jones, Nic, Waterfield, Michael D., Timms, John F.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01-05-2006; WILEY‐VCH Verlag
• Subjects: Electrophoresis, Gel, Two-Dimensional; Gene Deletion; Gene Expression Regulation; Hydrogen peroxide; Hydrogen Peroxide - metabolism; Hydrogen Peroxide - pharmacology; Mass Spectrometry; Metabolomics; Mitogen-Activated Protein Kinase Kinases - metabolism; Mitogen-Activated Protein Kinases - drug effects; Mitogen-Activated Protein Kinases - genetics; Mitogen-Activated Protein Kinases - metabolism; Oxidants - metabolism; Oxidants - pharmacology; Oxidative Stress - drug effects; Oxidative Stress - physiology; Protein Isoforms - drug effects; Protein Isoforms - metabolism; Protein Processing, Post-Translational; Proteomics; Redox stress; RNA, Messenger - biosynthesis; Schizosaccharomyces - drug effects; Schizosaccharomyces - genetics; Schizosaccharomyces - metabolism; Schizosaccharomyces pombe; Schizosaccharomyces pombe Proteins - drug effects; Schizosaccharomyces pombe Proteins - genetics; Schizosaccharomyces pombe Proteins - metabolism
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200500741

Using an integrated approach incorporating proteomics, metabolomics and published mRNA data, we have investigated the effects of hydrogen peroxide on wild type and a Sty1p‐deletion mutant of the fission yeast Schizosaccharomyces pombe. Differential protein expression analysis based on the modification of proteins with matched fluorescent labelling reagents (2‐D‐DIGE) is the foundation of the quantitative proteomics approach. This study identifies 260 differentially expressed protein isoforms from 2‐D‐DIGE gels using MALDI MS and reveals the complexity of the cellular response to oxidative stress and the dependency on the Sty1p stress‐activated protein kinase. We show the relationship between these protein changes and mRNA expression levels identified in a parallel whole genome study, and discuss the regulatory mechanisms involved in protecting cells against hydrogen peroxide and the involvement of Sty1p‐dependent stress‐activated protein kinase signalling. Metabolomic profiling of 29 intermediates using 1H NMR was also conducted alongside the protein analysis using the same sample sets, allowing examination of how the protein changes might affect the metabolic pathways and biological processes involved in the oxidative stress response. This combined analysis identifies a number of interlinked metabolic pathways that exhibit stress‐ and Sty1‐dependent patterns of regulation.