DOI: 10.1073/pnas.0406241102. Positive charge is an important feature of the C-terminal transport signal of the VirB/D4-translocated proteins of <i>Agrobacterium</i>

Positive charge is an important feature of the C-terminal transport signal of the VirB/D4-translocated proteins of Agrobacterium

10.1073/pnas.0406241102

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Several human pathogens and the plant pathogen Agrobacterium tumefaciens use a type IV secretion system for translocation of effector proteins into host cells. How effector proteins are selected for transport is unknown, but a C-terminal transport signal is present in the proteins translocated by the A. tumefaciens VirB/D4 type IV secretion system. We characterized this signal in the virulence protein VirF by alanine scanning and further site-directed mutagenesis. The Cre recombinase was used as a reporter to measure the translocation efficiency of Cre-Vir fusions from A. tumefaciens to Arabidopsis . The data unambiguously showed that positive charge is an essential characteristic of the C-terminal transport signal. We increased the sensitivity of this translocation assay by modifying the Cre-induced readout in host cells from kanamycin resistance to GFP expression. This improvement allowed us to detect translocation of the VirD2 relaxase protein in the absence of transferred DNA, showing that attachment to the transferred DNA is not essential for transport by the VirB/D4 system. We also found another translocated effector protein, namely the VirD5 protein encoded by the tumor-inducing plasmid. According to secondary structure predictions, the C termini of all VirB/D4-translocated proteins identified so far are unstructured; however, they contain a characteristic hydropathic profile. Based on sequence alignments and mutational analysis of VirF, we conclude that the C-terminal transport signal for recruitment and translocation of effector proteins by the A. tumefaciens VirB/D4 system is hydrophilic and has a net positive charge with a consensus motif of R-X(7)-R-X-R-X-R-X-X(n)>.

Bibliography

Vergunst, A. C., van Lier, M. C. M., den Dulk-Ras, A., Grosse StÃ¼ve, T. A., Ouwehand, A., & Hooykaas, P. J. J. (2005). Positive charge is an important feature of the C-terminal transport signal of the VirB/D4-translocated proteins of Agrobacterium. Proceedings of the National Academy of Sciences, 102(3), 832â837.

Authors 6

Annette C. Vergunst (first)
Miranda C. M. van Lier (additional)
Amke den Dulk-Ras (additional)
Thomas A. Grosse Stüve (additional)
Anette Ouwehand (additional)
Paul J. J. Hooykaas (additional)

References 39 Referenced 240

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Dates

Type	When
Created	20 years, 7 months ago (Jan. 11, 2005, 10:13 p.m.)
Deposited	3 years, 4 months ago (April 25, 2022, 11:02 p.m.)
Indexed	1 month, 1 week ago (July 24, 2025, 7:09 a.m.)
Issued	20 years, 7 months ago (Jan. 11, 2005)
Published	20 years, 7 months ago (Jan. 11, 2005)
Published Online	20 years, 7 months ago (Jan. 11, 2005)
Published Print	20 years, 7 months ago (Jan. 18, 2005)

Funders 0

None

BibTeX

@article{Vergunst_2005, title={Positive charge is an important feature of the C-terminal transport signal of the VirB/D4-translocated proteins of Agrobacterium}, volume={102}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0406241102}, DOI={10.1073/pnas.0406241102}, number={3}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Vergunst, Annette C. and van Lier, Miranda C. M. and den Dulk-Ras, Amke and Grosse Stüve, Thomas A. and Ouwehand, Anette and Hooykaas, Paul J. J.}, year={2005}, month=jan, pages={832–837} }

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