Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary; Premium Postdoctoral Research Program, Hungarian Academy of Sciences, Budapest, Hungary
Lili Anna Kenéz
Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
Sarolta Tóth
Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
Viktória Kiss
Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary
Ágnes Varga
Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
Tamás Csizmadia
Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary
Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary; Institute of Genetics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary
Two related multisubunit tethering complexes promote endolysosomal trafficking in all eukaryotes: Rab5-binding CORVET that was suggested to transform into Rab7-binding HOPS. We have previously identified miniCORVET, containing Drosophila Vps8 and three shared core proteins, which are required for endosome maturation upstream of HOPS in highly endocytic cells (Lőrincz et al., 2016a). Here, we show that Vps8 overexpression inhibits HOPS-dependent trafficking routes including late endosome maturation, autophagosome-lysosome fusion, crinophagy and lysosome-related organelle formation. Mechanistically, Vps8 overexpression abolishes the late endosomal localization of HOPS-specific Vps41/Lt and prevents HOPS assembly. Proper ratio of Vps8 to Vps41 is thus critical because Vps8 negatively regulates HOPS by outcompeting Vps41. Endosomal recruitment of miniCORVET- or HOPS-specific subunits requires proper complex assembly, and Vps8/miniCORVET is dispensable for autophagy, crinophagy and lysosomal biogenesis. These data together indicate the recruitment of these complexes to target membranes independent of each other in Drosophila, rather than their transformation during vesicle maturation.
