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2014 ; 10
(9
): 1588-602
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Role of VAMP3 and VAMP7 in the commitment of Yersinia pseudotuberculosis to
LC3-associated pathways involving single- or double-membrane vacuoles
#MMPMID25046114
Ligeon LA
; Moreau K
; Barois N
; Bongiovanni A
; Lacorre DA
; Werkmeister E
; Proux-Gillardeaux V
; Galli T
; Lafont F
Autophagy
2014[Sep]; 10
(9
): 1588-602
PMID25046114
show ga
Yersinia pseudotuberculosis can replicate inside macrophages by hijacking
autophagy and blocking autophagosome acidification. In bone marrow-derived
macrophages, the bacteria are mainly observed inside double-membrane vacuoles
positive for LC3, a hallmark of autophagy. Here, we address the question of the
membrane traffic during internalization of Yersinia investigating the role of
vesicle- associated membrane proteins (VAMPs). First, we show that as in
epithelial cells, Yersinia pseudotuberculosis replicates mainly in nonacidic
LC3-positive vacuoles. Second, in these cells, we unexpectedly found that VAMP3
localizes preferentially to Yersinia-containing vacuoles (YCVs) with single
membranes using correlative light-electron microscopy. Third, we reveal the
precise kinetics of VAMP3 and VAMP7 association with YCVs positive for LC3.
Fourth, we show that VAMP7 knockdown alters LC3's association with single-and
multimembrane-YCVs. Finally, in uninfected epithelial cells stimulated for
autophagy, VAMP3 overexpression and knockdown led respectively to a lower and
higher number of double-membrane, LC3-positive vesicles. Hence, our results
highlight the role that VAMPs play in selection of the pathways leading to
generation of ultrastructurally different LC3 compartments and pave the way for
determining the full set of docking and fusion proteins involved in Yersinia
pseudotuberculosis' intravesicular life cycle.