A parallel proteomic and metabolomic analysis of the hydrogen peroxide- and Sty1p-dependent stress response in Schizosaccharomyces pombe.
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.
Ludwig Institute for Cancer Research, University College London, Cruciform Building, London, UK.
Mark E
ME
Weeks
John
J
Sinclair
Amna
A
Butt
Yuen-Li
YL
Chung
Jessica L
JL
Worthington
Caroline R M
CR
Wilkinson
John
J
Griffiths
Nic
N
Jones
Michael D
MD
Waterfield
John F
JF
Timms
D015180:Electrophoresis, Gel, Two-Dimensional; D017353:Gene Deletion; D005786:Gene Expression Regulation; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology*; D013058:Mass Spectrometry; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D020033:Protein Isoforms / Q000187:drug effects / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D012333:RNA, Messenger / Q000096:biosynthesis; D012568:Schizosaccharomyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D016877:Oxidants; D020033:Protein Isoforms; D012333:RNA, Messenger; D029702:Schizosaccharomyces pombe Proteins; D006861:Hydrogen Peroxide; D020928:Mitogen-Activated Protein Kinases; C095409:sty1 protein, S pombe; D020929:Mitogen-Activated Protein Kinase Kinases