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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Hooykaas, Paul J. J. Vergunst, Annette C. Ouwehand, Anette Van Lier, Miranda C. M. Stüve, Thomas A. Grosse Den Dulk-ras, Amke |
| Description | Author Affiliation: Vergunst AC ( Clusius Laboratory, Institute of Biology, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.); |
| 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)>. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 3 |
| Volume Number | 102 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2005-01-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Membrane Transport Proteins Chemistry Protein Sorting Signals Rhizobium Pathogenicity Virulence Factors Arabidopsis Metabolism Bacterial Proteins Biological Transport Consensus Sequence Genes, Reporter Genetics Mutagenesis, Site-Directed Static Electricity Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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