RRC ID 54733
著者 Itai N, Shimazu T, Kimura T, Ibe I, Yamashita R, Kaburagi Y, Dohi T, Tonozuka T, Takao T, Nishikawa A.
タイトル The phosphorylation of sorting nexin 5 at serine 226 regulates retrograde transport and macropinocytosis.
ジャーナル PLoS One
Abstract Sorting nexin 5 (SNX5), a member of sorting nexin family, plays an important role in membrane trafficking, including the retrograde trafficking of the cation independent mannose 6-phosphate receptor (CI-M6PR) and macropinocytosis. Using ESI-LCMS/MS analysis, we confirmed that SNX5 serine 226 is phosphorylated. Since SNX5 forms heterodimers with SNX1 or SNX2, we examined the effect of phosphorylation at S226 on the heterodimer formations. Wild-type and mutants of SNX5, in which S226 was mutated to a glutamic acid or an alanine, were expressed in 8505C cells. In pull-down assays using SNX5 as bait, only the S226E mutant failed to precipitate both SNX1 and SNX2. Confocal microscopy data indicated that the wild type and S226A mutant were colocalized with SNX1 and SNX2 in endosomes, but the S226E was not. SNX5 and SNX6 support each other's functions and are involved with CI-M6PR retrograde trafficking. In SNX5 and SNX6 double knockdown cells, CI-M6PR was dispersed and colocalized with the endosomal marker EEA1. In a rescue experiment using SNX5 mutants, the S226A rescued CI-M6PR localization, similar to control cells, but S226E did not. Furthermore, the decrease in the uptake of dextran by macropinocytosis in SNX5 knockdown cells was recovered by the expression of rescue-wild type or S226A mutant, but not by the rescue-S226E mutant. These observations indicate that SNX5 constitutive phosphorylation that mimics the mutant S226E decreases the active SNX5 in these cells. The phosphorylation of SNX5 regulates the dimerization with SNX1 or SNX2, and this suggests that it controls membrane trafficking and protein sorting.
巻・号 13(11)
ページ e0207205
公開日 2018-11-12
DOI 10.1371/journal.pone.0207205
PII PONE-D-18-23603
PMID 30419003
PMC PMC6231649
MeSH Amino Acid Sequence Biological Transport / physiology* Cell Line, Tumor Dextrans / metabolism Endosomes / metabolism Humans Mutation Phosphorylation Pinocytosis / physiology* Protein Multimerization Receptor, IGF Type 2 / metabolism Sorting Nexins / genetics* Sorting Nexins / metabolism*
IF 2.776
引用数 2
リソース情報
ヒト・動物細胞 8505C(RCB2103) A549