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Defects in intracellular trafficking and endocytic/vacuolar acidification determine the efficiency of endocytotic DNA uptake in yeast.
RIECHERS, Sean-Patrick Hermann; Stahl, Ulf; Lang, Christine
2009In Journal of Cellular Biochemistry, 106 (2), p. 327-36
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Keywords :
Acids; Biological Transport; DNA, Fungal/metabolism; Endocytosis; Hydrogen-Ion Concentration; Mutation/genetics; Saccharomyces cerevisiae/genetics/metabolism; Transgenes/genetics; Vacuoles/metabolism
Abstract :
[en] 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 N+/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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
RIECHERS, Sean-Patrick Hermann  ;  Technische Universität Berlin, Department of Microbiology and Genetics, Institute for Biotechnology, Gustav-Meyer-Allee 25, D-13355 Berlin, Germany.
Stahl, Ulf
Lang, Christine
External co-authors :
yes
Language :
English
Title :
Defects in intracellular trafficking and endocytic/vacuolar acidification determine the efficiency of endocytotic DNA uptake in yeast.
Publication date :
2009
Journal title :
Journal of Cellular Biochemistry
ISSN :
0730-2312
eISSN :
1097-4644
Publisher :
John Wiley & Sons, Hoboken, United States - New Jersey
Volume :
106
Issue :
2
Pages :
327-36
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBilu :
since 12 September 2023

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