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Role of cholesterol and N-glycosylation in apical sorting of GPI- APs in polarized epithelial FRT cells

Abstract : Epithelial cells represent the ability to polarize with an apical and basolateral domains which differ markedly in proteins, lipid composition and therefore in function. This asymmetry reflects the ability of epithelial cells to sort newly synthesized proteins and lipid to either cell surface. While the signals responsible for basolateral targeting of the proteins have been clearly understood, the situation regarding the apical sorting of proteins is more obscure. We have previously shown that differently from basolateral GPI-APs oligomerization in the Golgi apparatus is necessary for apical sorting of Glycosylphosphatidylinositol- anchored proteins (GPI-APs). Interestingly this mechanism is conserved in two different kinds of epithelial cells, MDCK and FRT cells, which exhibits a difference in the sorting of GPI-APs. However the precise mechanism leading to this event is not understood. Our previous data demonstrated that simple addition of cholesterol to MDCK cells is necessary and sufficient to induce the oligomerization and apical sorting of a basolateral GPI-AP. Whereas, in this present study in FRT cells we showed that in contrast with MDCK cells cholesterol is not an active player in the regulation of GPI- APs apical sorting. In addition, we also showed that apical and basolateral GPI-APs are not segregated in the Golgi on the bases of the cholesterol content of the surrounding membrane environment. Furthermore, we demonstrated that N- glycosylation of the protein ectodomain is critical for oligomerization and apical sorting of GPI-APs. Our data indicates that at least two mechanisms depending either on cholesterol or on N-glycosylation exist to determine oligomerization in the Golgi and sorting to the apical membrane of GPI-APs.
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Submitted on : Saturday, March 24, 2012 - 3:42:11 PM
Last modification on : Thursday, April 7, 2022 - 10:10:30 AM
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Naga Sailaja Imjeti. Role of cholesterol and N-glycosylation in apical sorting of GPI- APs in polarized epithelial FRT cells. Agricultural sciences. Université Paris Sud - Paris XI, 2011. English. ⟨NNT : 2011PA112362⟩. ⟨tel-00682284⟩

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