title: LRRK2, but not pathogenic mutants, protects against H2O2 stress depending on mitochondrial function and endocytosis in a yeast model.
abstract: BACKGROUND
  Mutations in LRRK2 are the most common genetic cause of Parkinson's disease (PD). Studies in the yeast Saccharomyces cerevisiae have provided valuable insights into the mechanisms of cellular dysfunction associated with the expression of faulty PD genes.
  
  
  METHODS
  We developed a yeast model for full-length LRRK2 studies. We expressed wild-type (wt) LRRK2 and mutations and evaluated their role during oxidative stress conditions. The involvement of mitochondria was assessed by using rho-zero mutants and by evaluating reactive oxygen species (ROS) production and mitochondrial membrane potential by flow cytometry. The involvement of endocytosis was also studied by testing several endocytic mutants and by following the vacuolar delivery of the probe FM4-64.
  
  
  RESULTS
  Expression of LRRK2 in yeast was associated to increased hydrogen peroxide resistance. This phenotype, which was dependent on mitochondrial function, was not observed for PD-mutants G2019S and R1441C or in the absence of the kinase activity and the WD40 repeat domain. Expression of the pathogenic mutants stimulated ROS production and increased mitochondrial membrane potential. For the PD-mutants, but not for wild-type LRRK2, endocytic defects were also observed. Additionally, several endocytic proteins were required for LRRK2-mediated protection against hydrogen peroxide.
  
  
  CONCLUSIONS
  Our results indicate that LRRK2 confers cellular protection during oxidative stress depending on mitochondrial function and endocytosis.
  
  
  CONCLUSIONS
  Both the loss of capacity of LRRK2 pathogenic mutants to protect against oxidative stress and their enhancement of dysfunction may be important for the development of PD during the aging process.
authors.affiliation: REQUIMTE, Laboratório de Microbiologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal. Electronic address: clarafrancisco@gmail.com.
authors.forename: Clara
authors.initials: C
authors.lastname: Pereira
authors.affiliation: Cell Death Regulation Laboratory, MRC Toxicology Unit, Leicester, UK.
authors.forename: L
authors.initials: L
authors.lastname: Miguel Martins
authors.affiliation: REQUIMTE, Laboratório de Microbiologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal. Electronic address: lucilia.saraiva@ff.up.pt.
authors.forename: Lucília
authors.initials: L
authors.lastname: Saraiva
mesh_terms: D004705:Endocytosis; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008928:Mitochondria / Q000502:physiology*; D009154:Mutation; D018384:Oxidative Stress; D010300:Parkinson Disease / Q000209:etiology* / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D006861:Hydrogen Peroxide; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases
keywords: Cellular dysfunction; LRRK2; Oxidative stress; Parkinson's disease; Saccharomyces cerevisiae