title: Nuclear cytoplasmic trafficking of proteins is a major response of human fibroblasts to oxidative stress.
abstract: We have used a subcellular spatial razor approach based on LC-MS/MS-based proteomics with SILAC isotope labeling to determine changes in protein abundances in the nuclear and cytoplasmic compartments of human IMR90 fibroblasts subjected to mild oxidative stress. We show that response to mild tert-butyl hydrogen peroxide treatment includes redistribution between the nucleus and cytoplasm of numerous proteins not previously associated with oxidative stress. The 121 proteins with the most significant changes encompass proteins with known functions in a wide variety of subcellular locations and of cellular functional processes (transcription, signal transduction, autophagy, iron metabolism, TCA cycle, ATP synthesis) and are consistent with functional networks that are spatially dispersed across the cell. Both nuclear respiratory factor 2 and the proline regulatory axis appear to contribute to the cellular metabolic response. Proteins involved in iron metabolism or with iron/heme as a cofactor as well as mitochondrial proteins are prominent in the response. Evidence suggesting that nuclear import/export and vesicle-mediated protein transport contribute to the cellular response was obtained. We suggest that measurements of global changes in total cellular protein abundances need to be complemented with measurements of the dynamic subcellular spatial redistribution of proteins to obtain comprehensive pictures of cellular function.
authors.affiliation: Division of Medicine, Center for Nephrology, University College London , Royal Free Campus, Rowland Hill Street, London NW3 2PF, United Kingdom.
authors.forename: Noor O
authors.initials: NO
authors.lastname: Baqader
authors.forename: Marko
authors.initials: M
authors.lastname: Radulovic
authors.forename: Mark
authors.initials: M
authors.lastname: Crawford
authors.forename: Kai
authors.initials: K
authors.lastname: Stoeber
authors.forename: Jasminka
authors.initials: J
authors.lastname: Godovac-Zimmermann
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002853:Chromatography, Liquid; D003593:Cytoplasm / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D018384:Oxidative Stress; D021381:Protein Transport; D053719:Tandem Mass Spectrometry
keywords: DNA replication; SILAC; mass spectrometry; oxidative stress; peroxide; quantitative proteomics