Syntaxin 16 and syntaxin 5 are required for efficient retrograde transport of several exogenous and endogenous cargo proteins.
Retrograde transport allows proteins and lipids to leave the endocytic pathway to reach other intracellular compartments, such as trans-Golgi network (TGN)/Golgi membranes, the endoplasmic reticulum and, in some instances, the cytosol. Here, we have used RNA interference against the SNARE proteins syntaxin 5 and syntaxin 16, combined with recently developed quantitative trafficking assays, morphological approaches and cell intoxication analysis to show that these SNARE proteins are not only required for efficient retrograde transport of Shiga toxin, but also for that of an endogenous cargo protein - the mannose 6-phosphate receptor - and for the productive trafficking into cells of cholera toxin and ricin. We have found that the function of syntaxin 16 was specifically required for, and restricted to, the retrograde pathway. Strikingly, syntaxin 5 RNA interference protected cells particularly strongly against Shiga toxin. Since our trafficking analysis showed that apart from inhibiting retrograde endosome-to-TGN transport, the silencing of syntaxin 5 had no additional effect on Shiga toxin endocytosis or trafficking from TGN/Golgi membranes to the endoplasmic reticulum, we hypothesize that syntaxin 5 also has trafficking-independent functions. In summary, our data demonstrate that several cellular and exogenous cargo proteins use elements of the same SNARE machinery for efficient retrograde transport between early/recycling endosomes and TGN/Golgi membranes.
Amessou
Mohamed
M
Traffic and Signaling Laboratory, UMR144Curie/CNRS, Institut Curie, 26 rue d'Ulm, F-75248 Paris Cedex 05, France.
Fradagrada
Alexandre
A
Falguières
Thomas
T
Lord
J Michael
JM
Smith
Daniel C
DC
Roberts
Lynne M
LM
Lamaze
Christophe
C
Johannes
Ludger
L
D001483:Base Sequence; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D021381:Protein Transport; D050765:Qa-SNARE Proteins / Q000502:physiology*; D034622:RNA Interference; D051438:Syntaxin 16 / Q000502:physiology*
D017931:DNA Primers; D050765:Qa-SNARE Proteins; D051438:Syntaxin 16