Transgenic sensors reveal compartment-specific effects of aggregation-prone proteins on subcellular proteostasis during aging

Cell Rep Methods. 2024 Oct 21;4(10):100875. doi: 10.1016/j.crmeth.2024.100875. Epub 2024 Oct 8.

Abstract

Loss of proteostasis is a hallmark of aging that underlies many age-related diseases. Different cell compartments experience distinctive challenges in maintaining protein quality control, but how aging regulates subcellular proteostasis remains underexplored. Here, by targeting the misfolding-prone FlucDM luciferase to the cytoplasm, mitochondria, and nucleus, we established transgenic sensors to examine subcellular proteostasis in Drosophila. Analysis of detergent-insoluble and -soluble levels of compartment-targeted FlucDM variants indicates that thermal stress, cold shock, and pro-longevity inter-organ signaling differentially affect subcellular proteostasis during aging. Moreover, aggregation-prone proteins that cause different neurodegenerative diseases induce a diverse range of outcomes on FlucDM insolubility, suggesting that subcellular proteostasis is impaired in a disease-specific manner. Further analyses with FlucDM and mass spectrometry indicate that pathogenic tauV337M produces an unexpectedly complex regulation of solubility for different FlucDM variants and protein subsets. Altogether, compartment-targeted FlucDM sensors pinpoint a diverse modulation of subcellular proteostasis by aging regulators.

Keywords: CP: cell biology; CP: molecular biology; cell compartments; inter-tissue signaling; myokines; organelles; protein quality control; subcellular proteostasis; tools for aging research.

MeSH terms

  • Aging* / metabolism
  • Animals
  • Animals, Genetically Modified
  • Drosophila
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / metabolism
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Protein Aggregates
  • Proteostasis*

Substances

  • Protein Aggregates
  • Drosophila Proteins
  • Luciferases