Defects in intracellular trafficking and endocytic/vacuolar acidification determine the efficiency of endocytotic DNA uptake in yeast
✍ Scribed by Sean-Patrick Riechers; Ulf Stahl; Christine Lang
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 264 KB
- Volume
- 106
- Category
- Article
- ISSN
- 0730-2312
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✦ Synopsis
Abstract
The yeast Saccharomyces cerevisiae is a standard model system to study endocytosis. Here we describe the examination of a representative subset of deletion mutants to identify and locate steps in endocytic transport, endosomal/lysosomal acidification and in intracellular transport of hydrolases in non‐viral transfection processes. When transport in late endocytosis is inhibited, transfection efficiency is significantly enhanced. Similarly, transfection efficiency is enhanced when the pH‐value of the endosomal/vacuolar system is modified. Transfection efficiency is furthermore elevated when the Na^+^/K^+^ transport in the endosomal system is disturbed. Finally, we observe enhanced transfection efficiency in mutants disturbed in the CVT/autophagy pathway and in hydrolase transport to the vacuole. In summary, non‐viral transfection efficiency can be significantly increased by either (i) inhibiting the transport of endocytosed material before it enters the vacuole, or (ii) inducing a non‐natural pH‐value of the endosomal/vacuolar system, or (iii) slowing down degradative processes by inhibiting vacuolar hydrolases or the transport between Golgi and late endosome/vacuole. J. Cell. Biochem. 106: 327–336, 2009. © 2008 Wiley‐Liss, Inc.