Localization of GDP-mannose transporter in the Golgi requires retrieval to the endoplasmic reticulum depending on its cytoplasmic tail and coatomer.
The Saccharomyces cerevisiae GDP-mannose transporter (GMT) encoded by the essential gene VRG4/VIG4 is a member of the nucleotide-sugar transporter family in the Golgi apparatus. We examined GMT in the secretory mutant cells to investigate the mechanism of its localization in the Golgi. At the nonpermissive temperature, most GMT was found in the endoplasmic reticulum of sec23ts cells, which have defective COPII, and in the vacuole of sec21ts cells, which have defective COPI. The C-terminal hydrophilic peptide of GMT that is exposed to the cytosol binds to Ret2p, a subunit of the COPI coat. Mutant peptide derivatives that have lost a cluster of lysine in the vicinity of the transmembrane domain had reduced binding activity to Ret2p and the GMT with this sequence was delivered to the vacuole. Our results indicate that GMT escapes from delivery to the vacuole by recycling to the endoplasmic reticulum and retrieval requires the lysine-rich C-terminal tail that can bind to the COPI coat.
Abe
Masato
M
Department of Biotechnology, University of Tokyo, Yayoi, Bunkyo-Ku, Tokyo 113-8657, Japan.
Noda
Yoichi
Y
Adachi
Hiroyuki
H
Yoda
Koji
K
D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D020755:Coat Protein Complex I / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism
D002352:Carrier Proteins; D020755:Coat Protein Complex I; C428326:GDP-mannose transporter