title: Effect of Lon protease knockdown on mitochondrial function in HeLa cells.
abstract: ATP-dependent proteases are currently emerging as key regulators of mitochondrial functions. Among these proteolytic systems, Lon protease is involved in the control of selective protein turnover in the mitochondrial matrix. In the absence of Lon, yeast cells have been shown to accumulate electron-dense inclusion bodies in the matrix space, to loose integrity of mitochondrial genome and to be respiratory deficient. In order to address the role of Lon in mitochondrial functionality in human cells, we have set up a HeLa cell line stably transfected with a vector expressing a shRNA under the control of a promoter which is inducible with doxycycline. We have demonstrated that reduction of Lon protease results in a mild phenotype in this cell line in contrast with what have been observed in other cell types such as WI-38 fibroblasts. Nevertheless, deficiency in Lon protease led to an increase in ROS production and to an accumulation of carbonylated protein in the mitochondria. Our study suggests that Lon protease has a wide variety of targets and is likely to play different roles depending of the cell type.
authors.affiliation: UR4 - Vieillissement, Stress, Inflammation, Sorbonne Universités, UPMC Univ Paris 06, Université Pierre et Marie Curie, 4 Place Jussieu, 75252 Paris Cedex 05, France; Inserm, Hopital Robert Debré, 75019 Paris, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Bayot
authors.affiliation: UR4 - Vieillissement, Stress, Inflammation, Sorbonne Universités, UPMC Univ Paris 06, Université Pierre et Marie Curie, 4 Place Jussieu, 75252 Paris Cedex 05, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Gareil
authors.affiliation: Centre de recherche de Gif-sur-Yvette, FRC 3115, Centre de Génétique Moléculaire, CNRS, UPR3404, 91198 Gif-sur-Yvette Cedex, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Chavatte
authors.affiliation: UR4 - Vieillissement, Stress, Inflammation, Sorbonne Universités, UPMC Univ Paris 06, Université Pierre et Marie Curie, 4 Place Jussieu, 75252 Paris Cedex 05, France.
authors.forename: Marie-Paule
authors.initials: MP
authors.lastname: Hamon
authors.affiliation: Inserm, Institut Cochin, 75014 Paris, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: L'Hermitte-Stead
authors.affiliation: Institut de Biochimie et Génétique Cellulaires, UMR 5095, CNRS, Université Bordeaux 2, France.
authors.forename: Florian
authors.initials: F
authors.lastname: Beaumatin
authors.affiliation: Institut de Biochimie et Génétique Cellulaires, UMR 5095, CNRS, Université Bordeaux 2, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Priault
authors.affiliation: Inserm, Hopital Robert Debré, 75019 Paris, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Rustin
authors.affiliation: Inserm, Institut Cochin, 75014 Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Lombès
authors.affiliation: UR4 - Vieillissement, Stress, Inflammation, Sorbonne Universités, UPMC Univ Paris 06, Université Pierre et Marie Curie, 4 Place Jussieu, 75252 Paris Cedex 05, France. Electronic address: bertrand.friguet@snv.jussieu.fr.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Friguet
authors.affiliation: UR4 - Vieillissement, Stress, Inflammation, Sorbonne Universités, UPMC Univ Paris 06, Université Pierre et Marie Curie, 4 Place Jussieu, 75252 Paris Cedex 05, France.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Bulteau
mesh_terms: D002460:Cell Line; D004318:Doxycycline / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000187:drug effects; D049070:Protease La / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D050050:Protein Carbonylation / Q000187:drug effects; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D024101:Mitochondrial Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D049070:Protease La; D004318:Doxycycline
keywords: Cellular redox status; HeLa cells; Lon protease; Mitochondria; Protein oxidation