Rab11-mediated post-Golgi transport of the sialyltransferase ST3GAL4 suggests a new mechanism for regulating glycosylation.
Glycosylation, the most common posttranslational modification of proteins, is a stepwise process that relies on tight regulation of subcellular glycosyltransferase location to control the addition of each monosaccharide. Glycosyltransferases primarily reside and function in the endoplasmic reticulum (ER) and the Golgi apparatus; whether and how they traffic beyond the Golgi, how this trafficking is controlled, and how it impacts glycosylation remain unclear. Our previous work identified a connection between N-glycosylation and Rab11, a key player in the post-Golgi transport that connects recycling endosomes and other compartments. To learn more about the specific role of Rab11, we knocked down Rab11 in HeLa cells. Our findings indicate that Rab11 knockdown results in a dramatic enhancement in the sialylation of N-glycans. Structural analyses of glycans using lectins and LC-MS revealed that α2,3-sialylation is selectively enhanced, suggesting that an α2,3-sialyltransferase that catalyzes the sialyation of glycoproteins is activated or upregulated as the result of Rab11 knockdown. ST3GAL4 is the major α2,3-sialyltransferase that acts on N-glycans; we demonstrated that the localization of ST3GAL4, but not the levels of its mRNA, protein, or donor substrate, was altered by Rab11 depletion. In knockdown cells, ST3GAL4 is densely distributed in the trans-Golgi network, compared with the wider distribution in the Golgi and in other peripheral puncta in control cells, whereas the α2,6-sialyltransferase ST6GAL1 is predominantly localized to the Golgi regardless of Rab11 knockdown. This indicates that Rab11 may negatively regulate α2,3-sialylation by transporting ST3GAL4 to post-Golgi compartments (PGCs), which is a novel mechanism of glycosyltransferase regulation.
['Kitano|Masato|M|', 'Kizuka|Yasuhiko|Y|', 'Sobajima|Tomoaki|T|', 'Nakano|Miyako|M|', 'Nakajima|Kazuki|K|', 'Misaki|Ryo|R|', 'Itoyama|Saki|S|', 'Harada|Yoichiro|Y|', 'Harada|Akihiro|A|', 'Miyoshi|Eiji|E|', 'Taniguchi|Naoyuki|N|']
D000818:Animals D006031:Glycosylation D006056:Golgi Apparatus D006367:HeLa Cells D006801:Humans D021381:Protein Transport D051381:Rats D012799:Sialyltransferases D020691:rab GTP-Binding Proteins D021601:trans-Golgi Network
sialyltransferase ST3GAL4 glycosylation posttranslational modification glycosyltransferase Rab11 glycan α2,3-sialylation N-linked glycosylation membrane traffic