Effects of mutations in potential phosphorylation sites on transcytosis of FcRn.
The neonatal Fc receptor, FcRn, transports immunoglobulin G (IgG) across intestinal epithelial cells of suckling rats and mice from the lumenal surface to the serosal surface. In cell culture models FcRn transports IgG bidirectionally, but there are differences in the mechanisms of transport in the two directions. We investigated the effects of mutations in the cytoplasmic domain of FcRn on apical to basolateral and basolateral to apical transport of Fc across rat inner medullary collecting duct (IMCD) cells. Basolateral to apical transport did not depend upon determinants in the cytoplasmic domain. In contrast, an essentially tailless FcRn was markedly impaired in apical to basolateral transport. Using truncation and substitution mutants, we identified serine-313 and serine-319 as phosphorylation sites in the cytoplasmic domain of FcRn expressed in Rat1 fibroblasts. Mutations at Ser-319 did not affect transcytosis across IMCD cells. FcRn-S313A was impaired in apical to basolateral transcytosis to the same extent as tailless FcRn, whereas FcRn-S313D transported at wild-type levels. FcRn-S313A recycled more Fc to the apical medium than the wild-type receptor, suggesting that Ser-313 is required to allow FcRn to be diverted from an apical recycling pathway to a transcytotic pathway.
Rosenstiel Center for Basic Biomedical Sciences, W. M. Keck Institute for Cellular Visualization, and Biology Department, Brandeis University, Waltham, MA 02254-9110, USA.
K M
KM
McCarthy
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M
Lam
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L
Subramanian
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R
Shakya
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Z
Wu
E E
EE
Newton
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NE
Simister
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002478:Cells, Cultured; D004705:Endocytosis / Q000502:physiology; D015395:Histocompatibility Antigens Class I; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism; D016297:Mutagenesis, Site-Directed / Q000235:genetics*; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D011961:Receptors, Fc / Q000235:genetics* / Q000378:metabolism*; D012694:Serine / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D015395:Histocompatibility Antigens Class I; D011961:Receptors, Fc; D012694:Serine; C100688:Fc receptor, neonatal