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Vol. 9, Issue 1, 173-189, January 1998
Biozentrum of the University of Basel, CH-4056 Basel, Switzerland
Positively charged Nanogold (Nanoprobes, Stony Brook, NY) has been
developed as a new marker to follow the endocytic pathway in yeast.
Positively charged Nanogold binds extensively to the surface of yeast
spheroplasts and is internalized in an energy-dependent manner.
Internalization of gold is blocked in the end3 mutant. During a time course of incubation of yeast spheroplasts with positively charged Nanogold at 15°C, the gold was detected
sequentially in small vesicles, a peripheral, vesicular/tubular
compartment that we designate as an early endosome, a multivesicular
body corresponding to the late endosome near the vacuole, and in the vacuole. Experiments examining endocytosis in the sec18
mutant showed an accumulation of positively charged Nanogold in
approximately 30-50 nm diameter vesicles. These vesicles most likely
represent the primary endocytic vesicles as no other intermediates were detected in the mutant cells, and they correspond in size to the first
vesicles detected in wild-type spheroplasts at 15°C. These data lend
strong support to the idea that the internalization step of endocytosis
in yeast involves formation of small vesicles of uniform size from the
plasma membrane.
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