The carboxy‐terminus of VirE2 from Agrobacterium tumefaciens is required for its transport to host cells by the virB ‐encoded type IV transport system

The carboxy‐terminus of VirE2 from Agrobacterium tumefaciens is required for its transport to... Agrobacterium tumefaciens transfers DNA from the resident ‘tumour‐inducing’ (Ti) plasmid into plant cells, where it can be stably integrated into the plant genome, ultimately resulting in crown gall tumour formation. The mobilized DNA molecule is a single‐stranded intermediate with VirD2 covalently bound to its 5′ end. Successful transport of the transferred DNA (T‐DNA) and integration of the DNA into the genome requires that additional proteins be transported to the plant as well, including the single‐stranded (ss)DNA‐binding protein, VirE2. The transport of these two different substrates occurs as a result of the activities of a type IV secretion system encoded by the virB operon. Although the substrates have been identified, the mechanism of their transport remains unknown. In the experiments described here, a region in one of these substrates, VirE2, necessary for transport is identified. The addition of a C‐terminal FLAG epitope tag to VirE2, or the deletion of its C‐terminal 18 amino acids, renders it non‐functional in A. tumefaciens. However, transgenic plants expressing either of these virE2 genes respond to virE2 mutants of A. tumefaciens by forming wild‐type tumours. These results indicate that this region of VirE2 is necessary for the protein to be transported into the plant cells, but is not necessary for its function within the plant. Additionally, these studies demonstrate that mutant forms of VirE2 lacking this region do not disrupt the activities of the VirB transporter and support the hypothesis that VirE2 and the VirD2 T‐strand are transported independently, even when they co‐exist in the same cell. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Microbiology Wiley

The carboxy‐terminus of VirE2 from Agrobacterium tumefaciens is required for its transport to host cells by the virB ‐encoded type IV transport system

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Publisher
Wiley
Copyright
Copyright © 2001 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0950-382X
eISSN
1365-2958
D.O.I.
10.1046/j.1365-2958.2001.02582.x
Publisher site
See Article on Publisher Site

Abstract

Agrobacterium tumefaciens transfers DNA from the resident ‘tumour‐inducing’ (Ti) plasmid into plant cells, where it can be stably integrated into the plant genome, ultimately resulting in crown gall tumour formation. The mobilized DNA molecule is a single‐stranded intermediate with VirD2 covalently bound to its 5′ end. Successful transport of the transferred DNA (T‐DNA) and integration of the DNA into the genome requires that additional proteins be transported to the plant as well, including the single‐stranded (ss)DNA‐binding protein, VirE2. The transport of these two different substrates occurs as a result of the activities of a type IV secretion system encoded by the virB operon. Although the substrates have been identified, the mechanism of their transport remains unknown. In the experiments described here, a region in one of these substrates, VirE2, necessary for transport is identified. The addition of a C‐terminal FLAG epitope tag to VirE2, or the deletion of its C‐terminal 18 amino acids, renders it non‐functional in A. tumefaciens. However, transgenic plants expressing either of these virE2 genes respond to virE2 mutants of A. tumefaciens by forming wild‐type tumours. These results indicate that this region of VirE2 is necessary for the protein to be transported into the plant cells, but is not necessary for its function within the plant. Additionally, these studies demonstrate that mutant forms of VirE2 lacking this region do not disrupt the activities of the VirB transporter and support the hypothesis that VirE2 and the VirD2 T‐strand are transported independently, even when they co‐exist in the same cell.

Journal

Molecular MicrobiologyWiley

Published: Sep 1, 2001

References

  • Protein secretion by Gram‐negative bacterial ABC exporters – a review
    Binet, Binet; Létoffé, Létoffé; Ghigo, Ghigo; Delepelaire, Delepelaire; Wandersman, Wandersman
  • Nuclear localization of VirE2 protein in plant cells
    Citovsky, Citovsky; Zupan, Zupan; Warnick, Warnick; Zambryski, Zambryski
  • A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis
    O'Callaghan, O'Callaghan; Cazevieille, Cazevieille; Allardet‐Servent, Allardet‐Servent; Boschiroli, Boschiroli; Bourg, Bourg; Foulongne, Foulongne
  • Common ancestry between the IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens
    Pohlman, Pohlman; Genetti, Genetti; Winans, Winans
  • Intracellular multiplication and human macrophage killing by Legionella pneumophila are inhibited by conjugal components of IncQ plasmid RSF1010
    Segal, Segal; Shuman, Shuman

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