VPS29-VPS35 intermediate of retromer is stable and may be involved in the retromer complex assembly process.
Retromer is a complex of proteins that functions in the endosome-to-Golgi retrieval cargo transport pathway. VPS35 works as the central subunit of retromer to recognize the cargos and binds with VPS29 and VPS26 via distinct domains. We show that deficiency of VPS35 or VPS29 accompanies degradation of other subunits, whereas VPS26 deficiency had no effect on VPS29 and VPS35 levels. Although VPS35 forms VPS26-VPS35 and VPS29-VPS35 sub-complexes with similar efficiency in vitro, VPS26-VPS35 was more easily degradable by the ubiquitin-proteasome-system than VPS29-VPS35. These results indicate that VPS29 and VPS35 form a biologically stable sub-complex in vivo.
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Atsuhito
A
Fuse
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan; Department of Research and Therapeutics for Movement Disorders, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan. Electronic address: nohuruya@juntendo.ac.jp.
Norihiko
N
Furuya
Laboratory of Morphology and Image Analysis, Biomedical Research Center, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan; Department of Cellular and Molecular Neuropathology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Soichiro
S
Kakuta
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Aki
A
Inose
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Masumi
M
Sato
Laboratory of Morphology and Image Analysis, Biomedical Research Center, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan; Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Masato
M
Koike
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan.
Shinji
S
Saiki
Department of Neurology, Juntendo University Graduate School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan. Electronic address: nhattori@juntendo.ac.jp.
Nobutaka
N
Hattori
D015153:Blotting, Western; D011992:Endosomes / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics; D059748:Proteolysis; D034622:RNA Interference; D025801:Ubiquitin / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*; D021601:trans-Golgi Network / Q000378:metabolism* / Q000648:ultrastructure
D046912:Multiprotein Complexes; D025801:Ubiquitin; C498296:VPS26A protein, human; C419572:VPS29 protein, human; C419573:VPS35 protein, human; D033921:Vesicular Transport Proteins; D046988:Proteasome Endopeptidase Complex
Cation independent mannose-6-phosphate receptor; Proteasome; Retromer; SNXs; Ubiquitin; VPS26; VPS29; VPS35