Open AccessVisualization of VirE2 protein translocation by the A grobacterium type IV secretion system into host cells
Author(s) -
Sakalis Philippe A.,
Heusden G. Paul H.,
Hooykaas Paul J. J.
Publication year - 2014
Publication title -
microbiologyopen
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.881
H-Index - 36
ISSN - 2045-8827
DOI - 10.1002/mbo3.152
Subject(s) - bimolecular fluorescence complementation , green fluorescent protein , chromosomal translocation , agrobacterium , agrobacterium tumefaciens , secretion , virulence , biology , complementation , microbiology and biotechnology , plant cell , genetics , transformation (genetics) , yeast , gene , biochemistry , mutant
Type IV secretion systems (T4 SS ) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen A grobacterium tumefaciens uses a T4 SS to deliver a VirD2‐single stranded DNA complex as well as the virulence proteins VirD5, VirE2, VirE3, and VirF into host cells so that these become genetically transformed. Besides plant cells, yeast and fungi can efficiently be transformed by A grobacterium . Translocation of virulence proteins by the T4 SS has so far only been shown indirectly by genetic approaches. Here we report the direct visualization of VirE2 protein translocation by using bimolecular fluorescence complementation (Bi FC ) and Split GFP visualization strategies. To this end, we cocultivated A grobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part, either fused to VirE2 (for Bi FC ) or not (Split GFP ). Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation. Evidence was obtained that filament formation was due to the association of VirE2 with the microtubuli.