title: Polarized cell motility induces hydrogen peroxide to inhibit cofilin via cysteine oxidation.
abstract: Mesenchymal cell motility is driven by polarized actin polymerization [1]. Signals at the leading edge recruit actin polymerization machinery to promote membrane protrusion, while matrix adhesion generates tractive force to propel forward movement. To work effectively, cell motility is regulated by a complex network of signaling events that affect protein activity and localization. H2O2 has an important role as a diffusible second messenger [2], and mediates its effects through oxidation of cysteine thiols. One cell activity influenced by H2O2 is motility [3]. However, a lack of sensitive and H2O2-specific probes for measurements in live cells has not allowed for direct observation of H2O2 accumulation in migrating cells or protrusions. In addition, the identities of proteins oxidized by H2O2 that contribute to actin dynamics and cell motility have not been characterized. We now show, as determined by fluorescence lifetime imaging microscopy, that motile cells generate H2O2 at membranes and cell protrusions and that H2O2 inhibits cofilin activity through oxidation of cysteines 139 (C139) and 147 (C147). Molecular modeling suggests that C139 oxidation would sterically hinder actin association, while the increased negative charge of oxidized C147 would lead to electrostatic repulsion of the opposite negatively charged surface. Expression of oxidation-resistant cofilin impairs cell spreading, adhesion, and directional migration. These findings indicate that H2O2 production contributes to polarized cell motility through localized cofilin inhibition and that there are additional proteins oxidized during cell migration that might have similar roles.
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Jenifer M
authors.initials: JM
authors.lastname: Cameron
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Mads
authors.initials: M
authors.lastname: Gabrielsen
authors.affiliation: Division of Biomedical Engineering, School of Engineering, University of Glasgow, Glasgow G12 8LT, UK.
authors.forename: Ya Hua
authors.initials: YH
authors.lastname: Chim
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: June
authors.initials: J
authors.lastname: Munro
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Ewan J
authors.initials: EJ
authors.lastname: McGhee
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: David
authors.initials: D
authors.lastname: Sumpton
authors.affiliation: Cardiovascular Division, The Rayne Institute, St. Thomas' Hospital, King's College London, London SE1 7EH, UK.
authors.forename: Philip
authors.initials: P
authors.lastname: Eaton
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Kurt I
authors.initials: KI
authors.lastname: Anderson
authors.affiliation: Division of Biomedical Engineering, School of Engineering, University of Glasgow, Glasgow G12 8LT, UK.
authors.forename: Huabing
authors.initials: H
authors.lastname: Yin
authors.affiliation: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK. Electronic address: m.olson@beatson.gla.ac.uk.
authors.forename: Michael F
authors.initials: MF
authors.lastname: Olson
mesh_terms: D051339:Actin Depolymerizing Factors / Q000378:metabolism*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D010084:Oxidation-Reduction
chemical_list: D051339:Actin Depolymerizing Factors; D006861:Hydrogen Peroxide