Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells.

Autor: Sakalis PA; Institute of Biology, Leiden University, Sylviusweg 72, Leiden, 2333 BE, The Netherlands., van Heusden GP, Hooykaas PJ
Jazyk: angličtina
Zdroj: MicrobiologyOpen [Microbiologyopen] 2014 Feb; Vol. 3 (1), pp. 104-17. Date of Electronic Publication: 2013 Dec 27.
DOI: 10.1002/mbo3.152
Abstrakt: Type IV secretion systems (T4SS) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen Agrobacterium tumefaciens uses a T4SS 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 Agrobacterium. Translocation of virulence proteins by the T4SS 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 (BiFC) and Split GFP visualization strategies. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part, either fused to VirE2 (for BiFC) 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.
(© 2013 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
Externí odkaz: