title: A SPOPL/Cullin-3 ubiquitin ligase complex regulates endocytic trafficking by targeting EPS15 at endosomes.
abstract: Cullin-3 (CUL3)-based ubiquitin ligases regulate endosome maturation and trafficking of endocytic cargo to lysosomes in mammalian cells. Here, we report that these functions depend on SPOPL, a substrate-specific CUL3 adaptor. We find that SPOPL associates with endosomes and is required for both the formation of multivesicular bodies (MVBs) and the endocytic host cell entry of influenza A virus. In SPOPL-depleted cells, endosomes are enlarged and fail to acquire intraluminal vesicles (ILVs). We identify a critical substrate ubiquitinated by CUL3-SPOPL as EPS15, an endocytic adaptor that also associates with the ESCRT-0 complex members HRS and STAM on endosomes. Indeed, EPS15 is ubiquitinated in a SPOPL-dependent manner, and accumulates with HRS in cells lacking SPOPL. Together, our data indicates that a CUL3-SPOPL E3 ubiquitin ligase complex regulates endocytic trafficking and MVB formation by ubiquitinating and degrading EPS15 at endosomes, thereby influencing influenza A virus infection as well as degradation of EGFR and other EPS15 targets.
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Michaela
authors.initials: M
authors.lastname: Gschweitl
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Anna
authors.initials: A
authors.lastname: Ulbricht
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Barnes
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Radoslav I
authors.initials: RI
authors.lastname: Enchev
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Stoffel-Studer
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Meyer-Schaller
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Jatta
authors.initials: J
authors.lastname: Huotari
authors.affiliation: Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yamauchi
authors.affiliation: Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
authors.forename: Urs F
authors.initials: UF
authors.lastname: Greber
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Ari
authors.initials: A
authors.lastname: Helenius
authors.affiliation: Institute of Biochemistry, Department of Biology, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland.
authors.forename: Matthias
authors.identifier: http://orcid.org/0000-0002-2160-6824
authors.initials: M
authors.lastname: Peter
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D001692:Biological Transport; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002460:Cell Line; D044842:Cullin Proteins / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism*; D006801:Humans; D009980:Influenza A virus / Q000502:physiology; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C503645:CUL3 protein, human; D002135:Calcium-Binding Proteins; D044842:Cullin Proteins; C526966:Eps15 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; C000612949:SPOPL protein, human
keywords: CUL3; EGFR; EPS15; SPOPL; biochemistry; cell biology; endocytosis; human; influenza A virus