PMID: 11309206Apr 20, 2001Paper

Cholesterol requirement for cation-independent mannose 6-phosphate receptor exit from multivesicular late endosomes to the Golgi

Journal of Cell Science
I MiwakoM Ohashi

Abstract

The regulation of endocytic traffic of receptors has central importance in the fine tuning of cell activities. Here, we provide evidence that cholesterol is required for the exit of cation-independent mannose 6-phosphate receptor (CI-MPR) from the endosomal carrier vesicle/multivesicular bodies (ECV/MVBs) to the Golgi. A previously established Chinese hamster ovary cell mutant, LEX2, exhibits arrested ECV/MVBs in which CI-MPR and lysosomal glycoprotein-B (lgp-B) are accumulated. The abnormal accumulation of CI-MPR within the ECV/MVBs in LEX2 cells was corrected in a post-translational manner by the supplementation of medium with cholesterol. Furthermore, it was shown that, by expression cloning using LEX2 mutant, the introduction of the NAD(P)H steroid dehydrogenase-like protein, an enzyme involved in the later stage of cholesterol biosynthesis, allows the exit of CI-MPR from the MVBs to the Golgi and reduces the number of arrested ECV/MVBs in LEX2 cells. The recovery of the exit transport of CI-MPR from the ECV/MVBs was associated with the restoration of the normal cellular free cholesterol level and segregation between CI-MPR and lgp-B, both of which had been localized at the internal small vesicles of the arrested ECV/MVBs. ...Continue Reading

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