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The V-ATPase complex component RNAseK is required for lysosomal hydrolase delivery and autophagosome degradation

Author

Listed:
  • Agata N. Makar

    (University of Edinburgh, Crewe Road South)

  • Alina Boraman

    (University of Edinburgh, Crewe Road South)

  • Peter Mosen

    (University of Bonn)

  • Joanne E. Simpson

    (University of Edinburgh, Crewe Road South)

  • Jair Marques

    (University of Edinburgh, Crewe Road South)

  • Tim Michelberger

    (University of Edinburgh, Crewe Road South)

  • Stuart Aitken

    (Crewe Road South, University of Edinburgh)

  • Ann P. Wheeler

    (Crewe Road South, University of Edinburgh)

  • Dominic Winter

    (University of Bonn)

  • Alex Kriegsheim

    (University of Edinburgh, Crewe Road South)

  • Noor Gammoh

    (University of Edinburgh, Crewe Road South)

Abstract

Autophagy is a finely orchestrated process required for the lysosomal degradation of cytosolic components. The final degradation step is essential for clearing autophagic cargo and recycling macromolecules. Using a CRISPR/Cas9-based screen, we identify RNAseK, a highly conserved transmembrane protein, as a regulator of autophagosome degradation. Analyses of RNAseK knockout cells reveal that, while autophagosome maturation is intact, cargo degradation is severely disrupted. Importantly, lysosomal protease activity and acidification remain intact in the absence of RNAseK suggesting a specificity to autolysosome degradation. Analyses of lysosome fractions show reduced levels of a subset of hydrolases in the absence of RNAseK. Of these, the knockdown of PLD3 leads to a defect in autophagosome clearance. Furthermore, the lysosomal fraction of RNAseK-depleted cells exhibits an accumulation of the ESCRT-III complex component, VPS4a, which is required for the lysosomal targeting of PLD3. Altogether, here we identify a lysosomal hydrolase delivery pathway required for efficient autolysosome degradation.

Suggested Citation

  • Agata N. Makar & Alina Boraman & Peter Mosen & Joanne E. Simpson & Jair Marques & Tim Michelberger & Stuart Aitken & Ann P. Wheeler & Dominic Winter & Alex Kriegsheim & Noor Gammoh, 2024. "The V-ATPase complex component RNAseK is required for lysosomal hydrolase delivery and autophagosome degradation," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
  • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-52049-3
    DOI: 10.1038/s41467-024-52049-3
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