Glucose-induced ubiquitylation and endocytosis of the yeast Jen1 transporter: role of lysine 63-linked ubiquitin chains.
Protein ubiquitylation is essential for many events linked to intracellular protein trafficking. Despite the significance of this process, the molecular mechanisms that govern the regulation of ubiquitylation remain largely unknown. Plasma membrane transporters are subjected to tightly regulated endocytosis, and ubiquitylation is a key signal at several stages of the endocytic pathway. The yeast monocarboxylate transporter Jen1 displays glucose-regulated endocytosis. We show here that casein kinase 1-dependent phosphorylation and HECT-ubiquitin ligase Rsp5-dependent ubiquitylation are required for Jen1 endocytosis. Ubiquitylation and endocytosis of Jen1 are induced within minutes in response to glucose addition. Jen1 is modified at the cell surface by oligo-ubiquitylation with ubiquitin-Lys(63) linked chain(s), and Jen1-Lys(338) is one of the target residues. Ubiquitin-Lys(63)-linked chain(s) are also required directly or indirectly to sort Jen1 into multivesicular bodies. Jen1 is one of the few examples for which ubiquitin-Lys(63)-linked chain(s) was shown to be required for correct trafficking at two stages of endocytosis: endocytic internalization and sorting at multivesicular bodies.
Paiva
Sandra
S
Department of Biology, Molecular and Environmental Biology Centre, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal. spaiva@bio.uminho.pt
Vieira
Neide
N
Nondier
Isabelle
I
Haguenauer-Tsapis
Rosine
R
Casal
Margarida
M
Urban-Grimal
Danièle
D
D019943:Amino Acid Substitution; D001120:Arginine / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D015153:Blotting, Western; D047389:Casein Kinase I / Q000235:genetics / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects*; D056827:Endosomal Sorting Complexes Required for Transport; D005947:Glucose / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D027501:Monocarboxylic Acid Transporters / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism; D043743:Ubiquitin-Protein Ligase Complexes / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination / Q000187:drug effects
D056827:Endosomal Sorting Complexes Required for Transport; C118697:JEN1 protein, S cerevisiae; D027501:Monocarboxylic Acid Transporters; D029701:Saccharomyces cerevisiae Proteins; D027981:Symporters; D025801:Ubiquitin; D049452:Green Fluorescent Proteins; D001120:Arginine; D043743:Ubiquitin-Protein Ligase Complexes; D047389:Casein Kinase I; C479664:YCK1 protein, S cerevisiae; C469287:RSP5 protein, S cerevisiae; D005947:Glucose; D008239:Lysine