Biochemical Journal

Research article

Loss of TRPML1 promotes production of reactive oxygen species: is oxidative damage a factor in mucolipidosis type IV?

Jessica Coblentz, Claudette St. Croix, Kirill Kiselyov


TRPML1 (transient receptor potential mucolipin 1) is a lysosomal ion channel permeable to cations, including Fe2+. Mutations in MCOLN1, the gene coding for TRPML1, cause the LSD (lysosomal storage disease) MLIV (mucolipidosis type IV). The role of TRPML1 in the cell is disputed and the mechanisms of cell deterioration in MLIV are unclear. The demonstration of Fe2+ buildup in MLIV cells raised the possibility that TRPML1 dissipates lysosomal Fe2+ and prevents its accumulation. Since Fe2+ catalyses the production of ROS (reactive oxygen species), we set out to test whether or not the loss of TRPML1 promotes ROS production by Fe2+ trapped in lysosomes. Our data show that RPE1 (retinal pigmented epithelial 1) cells develop a punctate mitochondrial phenotype within 48 h of siRNA-induced TRPML1-KD (knockdown). This mitochondrial fragmentation was aggravated by Fe2+ exposure, but was reversed by incubation with the ROS chelator α-Toc (α-tocopherol). The exposure of TRPML1-KD cells to Fe2+ led to loss of ΔΨm (mitochondrial membrane potential), ROS buildup, lipid peroxidation and increased transcription of genes responsive to cytotoxic oxidative stress in TRPML1-KD cells. These data suggest that TRPML1 redistributes Fe2+ between the lysosomes and the cytoplasm. Fe2+ buildup caused by TRPML1 loss potentiates ROS production and leads to mitochondrial deterioration. Beyond suggesting a new model for MLIV pathogenesis, these data show that TRPML1's role in the cell extends outside lysosomes.

  • iron
  • lysosomal storage disease
  • mitochondrion
  • reactive oxygen species
  • transient receptor potential (TRP) channel

Abbreviations: AR, aspect ratio; BODIPY®, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; CCCP, carbonyl cyanide m-chlorophenylhydrazone; DMEM, Dulbecco’s modified Eagle’s medium; FF, form factor; HMOX1, haem oxygenase-1; JC-1, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide; KD, knockdown; LSD, lysosomal storage disease; 3-MA, 3-methyladenine; MLIV, mucolipidosis type IV; Nrf-2, nuclear factor erythroid 2-related factor 2; qPCR, quantitative real-time PCR; ROS, reactive oxygen species; RPE1, retinal pigmented epithelial 1; TBHP, t-butyl hydroperoxide; α-Toc, α-tocopherol; TRPML, transient receptor potential mucolipin; ΔΨm, mitochondrial membrane potential

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