title: Proteome alteration in oxidative stress-sensitive methionine sulfoxide reductase-silenced HEK293 cells.
abstract: Methionine sulfoxide reductases (Msr's) are key enzymes proficient in catalyzing the reduction of oxidized methionines. This reductive trait is essential to maintaining cellular redox homeostasis from bacteria to mammals and is also regarded as a potential mechanism to regulate protein activities and signaling pathways, considering the inactivating effects that can be induced by methionine oxidation. In this study, we have generated stable human embryonic kidney HEK293 clones with an altered Msr system by silencing the expression of the main Msr elements-MsrA, MsrB1, or MsrB2. The isolated clones--the single mutants MsrA, MsrB1, and MsrB2 and double mutant MsrA/B1-show a reduced Msr activity and an exacerbated sensitivity toward oxidative stress. A two-dimensional difference in-gel electrophoresis analysis was performed on the Msr-silenced cells grown under basal conditions or submitted to oxidative stress. This proteomic analysis revealed that the disruption of the Msr system mainly affects proteins with redox, cytoskeletal or protein synthesis, and maintenance roles. Interestingly, most of the proteins found altered in the Msr mutants were also identified as potential Msr substrates and have been associated with redox or aging processes in previous studies. This study, through an extensive analysis of Msr-inhibited mutants, offers valuable input on the cellular network of a crucial maintenance system such as methionine sulfoxide reductases.
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France. Electronic address: nicolasugarte@yahoo.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ugarte
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Ladouce
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Radjei
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Gareil
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Friguet
authors.affiliation: Laboratoire de Biologie Cellulaire du Vieillissement, UR4, IFR83, Université Pierre et Marie Curie-Paris 6, 75005 Paris, France. Electronic address: isabelle.petropoulos@snv.jussieu.fr.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Petropoulos
mesh_terms: D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D051150:Methionine Sulfoxide Reductases / Q000235:genetics* / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D060066:Protein Interaction Maps; D020543:Proteome / Q000378:metabolism*; D034622:RNA Interference; D013654:Taurine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D016877:Oxidants; D020543:Proteome; D013654:Taurine; C043410:N-chlorotaurine; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase
keywords: 2D-DIGE; Adaptive response; Free radicals; HEK; Methionine sulfoxide reductases; Msr; Oxidative stress; Oxidized protein repair; Proteomics; RNA interference; RNA silencing; RNAi; ROS; TnCl; human embryonic kidney cell line; methionine sulfoxide reductase; miRNA; microRNA; reactive oxygen species; taurine chloramine.; two-dimensional difference in-gel electrophoresis