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Vol. 13, Issue 8, 2664-2680, August 2002
and
*Biozentrum, University of Basel, CH-4056 Basel, Switzerland; and
Sterols are essential factors for endocytosis in animals and yeast.
To investigate the sterol structural requirements for yeast
endocytosis, we created a variety of ergInstitut für Biochemie, Technische
Universität, A-8010 Graz, Austria
mutants,
each accumulating a distinct set of sterols different from ergosterol. Mutant erg2erg6 and
erg3erg6 cells exhibit a strong
internalization defect of the 
-factor receptor (Ste2p). Specific
sterol structures are necessary for pheromone-dependent receptor
hyperphosphorylation, a prerequisite for internalization. The lack of
phosphorylation is not due to a defect in Ste2p localization or in
ligand-receptor interaction. Contrary to most known endocytic factors,
sterols seem to function in internalization independently of actin.
Furthermore, sterol structures are required at a postinternalization
step of endocytosis. erg cells were able to take up the
membrane marker FM4-64, but exhibited defects in FM4-64 movement
through endosomal compartments to the vacuole. Therefore, there are at
least two roles for sterols in endocytosis. Based on sterol analysis,
the sterol structural requirements for these two processes were
different, suggesting that sterols may have distinct functions at
different places in the endocytic pathway. Interestingly, sterol
structures unable to support endocytosis allowed transport of the
glycosylphosphatidylinositol-anchored protein Gas1p from
the endoplasmic reticulum to Golgi compartment.
Current address: Sainsbury Laboratory, John Innes
Center, Colney Lane, Norwich NR4 7UH, England.
§
Corresponding author. E-mail address:
howard.riezman{at}unibas.ch.
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