Related Experiment Video
Updated: May 2, 2026
Measurements of Physiological Stress Responses in C. Elegans
Published on: May 21, 2020
LRRK2, but not pathogenic mutants, protects against H2O2 stress depending on mitochondrial function and endocytosis
Clara Pereira1, L Miguel Martins2, Lucília Saraiva1
1REQUIMTE, Laboratório de Microbiologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal.
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.
General Significance:
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.
Insights
Mutations in Leucine-rich repeat kinase 2 (LRRK2) cause Parkinson's disease (PD). Our yeast model shows LRRK2 protects cells from oxidative stress via mitochondria and endocytosis, but PD-mutants lose this protective function.
Area of Science:
- Cell Biology
- Neuroscience
- Genetics
Background:
- Mutations in Leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of Parkinson's disease (PD).
- Studies in yeast (Saccharomyces cerevisiae) offer insights into cellular dysfunction mechanisms in PD.
Purpose of the Study:
- To develop and utilize a yeast model for studying full-length LRRK2, including wild-type and pathogenic mutations.
- To investigate the role of LRRK2 in cellular response to oxidative stress, focusing on mitochondrial function and endocytosis.
Main Methods:
- Expressed wild-type (wt) and mutant LRRK2 in yeast.
- Assessed cellular response to oxidative stress (hydrogen peroxide).
- Evaluated mitochondrial involvement using rho-zero mutants, reactive oxygen species (ROS) production, and mitochondrial membrane potential via flow cytometry.
- Studied endocytosis by testing endocytic mutants and tracking FM4-64 dye delivery to vacuoles.
Main Results:
- Yeast expressing LRRK2 showed increased hydrogen peroxide resistance, dependent on mitochondrial function.
- This protective effect was lost for PD-associated mutants (G2019S, R1441C) and in the absence of kinase activity or the WD40 domain.
- Pathogenic LRRK2 mutants increased ROS production and mitochondrial membrane potential.
- Endocytic defects were observed in PD-mutants, and endocytic proteins were necessary for LRRK2-mediated protection.
Conclusions:
- LRRK2 confers cellular protection against oxidative stress, a process dependent on mitochondrial function and endocytosis.
- The loss of protective capacity and enhanced dysfunction of LRRK2 pathogenic mutants may contribute to Parkinson's disease development during aging.
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