Lysosomal putative RNA transporter SIDT2 mediates direct uptake of RNA by lysosomes

Autophagy. 2016;12(3):565-78. doi: 10.1080/15548627.2016.1145325.

Abstract

Lysosomes are thought to be the major intracellular compartment for the degradation of macromolecules. We recently identified a novel type of autophagy, RNautophagy, where RNA is directly taken up by lysosomes in an ATP-dependent manner and degraded. However, the mechanism of RNA translocation across the lysosomal membrane and the physiological role of RNautophagy remain unclear. In the present study, we performed gain- and loss-of-function studies with isolated lysosomes, and found that SIDT2 (SID1 transmembrane family, member 2), an ortholog of the Caenorhabditis elegans putative RNA transporter SID-1 (systemic RNA interference deficient-1), mediates RNA translocation during RNautophagy. We also observed that SIDT2 is a transmembrane protein, which predominantly localizes to lysosomes. Strikingly, knockdown of Sidt2 inhibited up to ˜50% of total RNA degradation at the cellular level, independently of macroautophagy. Moreover, we showed that this impairment is mainly due to inhibition of lysosomal RNA degradation, strongly suggesting that RNautophagy plays a significant role in constitutive cellular RNA degradation. Our results provide a novel insight into the mechanisms of RNA metabolism, intracellular RNA transport, and atypical types of autophagy.

Keywords: RNA; RNautophagy; autophagy; lysosome; membrane protein.

MeSH terms

  • Animals
  • Autophagy
  • Gene Knockdown Techniques
  • HeLa Cells
  • Humans
  • Lysosomal-Associated Membrane Protein 2 / metabolism
  • Lysosomes / metabolism*
  • Membrane Proteins / metabolism*
  • Mice
  • Nucleotide Transport Proteins / metabolism*
  • Protein Binding
  • RNA / metabolism*
  • RNA Stability
  • RNA Transport*

Substances

  • Lysosomal-Associated Membrane Protein 2
  • Membrane Proteins
  • Nucleotide Transport Proteins
  • SIDT2 protein, human
  • Sidt2 protein, mouse
  • RNA

Grants and funding

This work was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (24680038 and 26111526 to T.K.), Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (to K.W.), and Grants-in-Aid for Scientific Research of the Ministry of Health, Labor and Welfare, Japan (to K.W.).