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. Author manuscript; available in PMC: 2014 May 12.
Published in final edited form as: Cell Rep. 2014 Feb 27;6(5):878–891. doi: 10.1016/j.celrep.2014.01.040

Figure 6. Disruption of the Golgi Architecture through ARF1-Dependent Membrane Tethering.

Figure 6

(A) CLEM shows the vesicular ultrastructure of the Golgi after microinjection with EspG, which remains constrained to areas positive for EspG signal.

(B) Identification of golgin GMAP-210 as an ARF1-dependent membrane tether involved in the EspG mechanism of Golgi disassembly by linking vesicles to membranes and preventing their transport.

(C) Fluorescent micrographs showing the distribution of GMAP-210 relative to the Golgi (GM130). GMAP-210 is retained on fragmented membranes following disassembly of the Golgi by EspG (arrowheads), in contrast to when ARF1 is removed from the membranes by BFA treatment.

(D) Pull-down experiment showing the binding of EspG to GMAP-210/ARF1 complex.

(E) Cartoon schematic illustrating the design of an ARF1-dependent, membrane-tethering chimera using a vesicle-binding motif linked to an ARF1-GTP-binding domain of GGA1. Our model of EspG function predicts vesicle-to-membrane linkage that is driven by the presence of GTP-ARF1 on membranes.

(F) Overexpression of the ARF1-dependent, membrane-tethering chimera is sufficient to disrupt the Golgi architecture. Transfected cells are marked with an asterisk.

(G) Control experiments show no impact on Golgi organization due to overexpression of either a vesicle-binding motif or an ARF1-GTP-binding domain alone in the absence of a direct tether.

See also Figure S5.