Previous Article | Next Article 
Journal of Bacteriology, August 1999, p. 4790-4797, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The Avr (Effector) Proteins HrmA (HopPsyA) and
AvrPto Are Secreted in Culture from Pseudomonas syringae
Pathovars via the Hrp (Type III) Protein Secretion System in a
Temperature- and pH-Sensitive Manner
Karin
van
Dijk,1
Derrick E.
Fouts,2
Amos H.
Rehm,2
Angela R.
Hill,1
Alan
Collmer,2 and
James R.
Alfano1,*
Department of Biological Sciences, University
of Nevada, Las Vegas, Nevada 89154-4004,1
and Department of Plant Pathology, Cornell University,
Ithaca, New York 14583-42032
Received 12 April 1999/Accepted 4 June 1999
We present here data showing that the Avr proteins HrmA and AvrPto
are secreted in culture via the native Hrp pathways from Pseudomonas syringae pathovars that produce these proteins.
Moreover, their secretion is strongly affected by the temperature and
pH of the culture medium. Both HrmA and AvrPto were secreted at their highest amounts when the temperature was between 18 and 22°C and when
the culture medium was pH 6.0. In contrast, temperature did not affect
the secretion of HrpZ. pH did affect HrpZ secretion, but not as
strongly as it affected the secretion of HrmA. This finding suggests
that there are at least two classes of proteins that travel the
P. syringae pathway: putative secretion system accessory
proteins, such as HrpZ, which are readily secreted in culture; and
effector proteins, such as HrmA and AvrPto, which apparently are
delivered inside plant cells and are detected in lower amounts in
culture supernatants under the appropriate conditions. Because HrmA was
shown to be a Hrp-secreted protein, we have changed the name of
hrmA to hopPsyA to reflect that it encodes a
Hrp outer protein from P. syringae pv. syringae. The
functional P. syringae Hrp cluster encoded by cosmid pHIR11
conferred upon P. fluorescens but not Escherichia
coli the ability to secrete HopPsyA in culture. The use of these
optimized conditions should facilitate the identification of additional
proteins traveling the Hrp pathway and the signals that regulate this
protein traffic.
*
Corresponding author. Mailing address: Department of
Biological Sciences, University of Nevada, Las Vegas, 4505 Maryland
Parkway, Box 454004, Las Vegas, NV 89154-4004. Phone: (702) 895-4420. Fax: (702) 895-3956. E-mail: alfanoj{at}nevada.edu.
Journal of Bacteriology, August 1999, p. 4790-4797, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Dawson, J. E., Seckute, J., De, S., Schueler, S. A., Oswald, A. B., Nicholson, L. K.
(2009). Elucidation of a pH-folding switch in the Pseudomonas syringae effector protein AvrPto. Proc. Natl. Acad. Sci. USA
106: 8543-8548
[Abstract]
[Full Text]
-
Morello, J. E., Collmer, A.
(2009). Pseudomonas syringae HrpP Is a Type III Secretion Substrate Specificity Switch Domain Protein That Is Translocated into Plant Cells but Functions Atypically for a Substrate-Switching Protein. J. Bacteriol.
191: 3120-3131
[Abstract]
[Full Text]
-
Griebel, T., Zeier, J.
(2008). Light Regulation and Daytime Dependency of Inducible Plant Defenses in Arabidopsis: Phytochrome Signaling Controls Systemic Acquired Resistance Rather Than Local Defense. Plant Physiol.
147: 790-801
[Abstract]
[Full Text]
-
Jones, A. M.E., Thomas, V., Bennett, M. H., Mansfield, J., Grant, M.
(2006). Modifications to the Arabidopsis Defense Proteome Occur Prior to Significant Transcriptional Change in Response to Inoculation with Pseudomonas syringae. Plant Physiol.
142: 1603-1620
[Abstract]
[Full Text]
-
Fu, Z. Q., Guo, M., Alfano, J. R.
(2006). Pseudomonas syringae HrpJ Is a Type III Secreted Protein That Is Required for Plant Pathogenesis, Injection of Effectors, and Secretion of the HrpZ1 Harpin.. J. Bacteriol.
188: 6060-6069
[Abstract]
[Full Text]
-
Brencic, A., Winans, S. C.
(2005). Detection of and Response to Signals Involved in Host-Microbe Interactions by Plant-Associated Bacteria. Microbiol. Mol. Biol. Rev.
69: 155-194
[Abstract]
[Full Text]
-
Smirnova, A. V., Ullrich, M. S.
(2004). Topological and deletion analysis of CorS, a Pseudomonas syringae sensor kinase. Microbiology
150: 2715-2726
[Abstract]
[Full Text]
-
Bretz, J. R., Hutcheson, S. W.
(2004). Role of Type III Effector Secretion during Bacterial Pathogenesis in Another Kingdom. Infect. Immun.
72: 3697-3705
[Full Text]
-
Wehling, M. D., Guo, M., Fu, Z. Q., Alfano, J. R.
(2004). The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone. J. Bacteriol.
186: 3621-3630
[Abstract]
[Full Text]
-
Schechter, L. M., Roberts, K. A., Jamir, Y., Alfano, J. R., Collmer, A.
(2004). Pseudomonas syringae Type III Secretion System Targeting Signals and Novel Effectors Studied with a Cya Translocation Reporter. J. Bacteriol.
186: 543-555
[Abstract]
[Full Text]
-
Keith, R. C., Keith, L. M. W., Hernandez-Guzman, G., Uppalapati, S. R., Bender, C. L.
(2003). Alginate gene expression by Pseudomonas syringae pv. tomato DC3000 in host and non-host plants. Microbiology
149: 1127-1138
[Abstract]
[Full Text]
-
Deng, W.-L., Rehm, A. H., Charkowski, A. O., Rojas, C. M., Collmer, A.
(2003). Pseudomonas syringae Exchangeable Effector Loci: Sequence Diversity in Representative Pathovars and Virulence Function in P. syringae pv. syringae B728a. J. Bacteriol.
185: 2592-2602
[Abstract]
[Full Text]
-
Casper-Lindley, C., Dahlbeck, D., Clark, E. T., Staskawicz, B. J.
(2002). Direct biochemical evidence for type III secretion-dependent translocation of the AvrBs2 effector protein into plant cells. Proc. Natl. Acad. Sci. USA
99: 8336-8341
[Abstract]
[Full Text]
-
Petnicki-Ocwieja, T., Schneider, D. J., Tam, V. C., Chancey, S. T., Shan, L., Jamir, Y., Schechter, L. M., Janes, M. D., Buell, C. R., Tang, X., Collmer, A., Alfano, J. R.
(2002). Genomewide identification of proteins secreted by the Hrp type III protein secretion system of Pseudomonas syringae pv. tomato DC3000. Proc. Natl. Acad. Sci. USA
99: 7652-7657
[Abstract]
[Full Text]
-
Guttman, D. S., Vinatzer, B. A., Sarkar, S. F., Ranall, M. V., Kettler, G., Greenberg, J. T.
(2002). A Functional Screen for the Type III (Hrp) Secretome of the Plant Pathogen Pseudomonas syringae. Science
295: 1722-1726
[Abstract]
[Full Text]
-
Fouts, D. E., Abramovitch, R. B., Alfano, J. R., Baldo, A. M., Buell, C. R., Cartinhour, S., Chatterjee, A. K., D'Ascenzo, M., Gwinn, M. L., Lazarowitz, S. G., Lin, N.-C., Martin, G. B., Rehm, A. H., Schneider, D. J., van Dijk, K., Tang, X., Collmer, A.
(2002). Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor. Proc. Natl. Acad. Sci. USA
99: 2275-2280
[Abstract]
[Full Text]
-
Hendrickson, E. L., Plotnikova, J., Mahajan-Miklos, S., Rahme, L. G., Ausubel, F. M.
(2001). Differential Roles of the Pseudomonas aeruginosa PA14 rpoN Gene in Pathogenicity in Plants, Nematodes, Insects, and Mice. J. Bacteriol.
183: 7126-7134
[Abstract]
[Full Text]
-
Hutcheson, S. W., Bretz, J., Sussan, T., Jin, S., Pak, K.
(2001). Enhancer-Binding Proteins HrpR and HrpS Interact To Regulate hrp-Encoded Type III Protein Secretion in Pseudomonas syringae Strains. J. Bacteriol.
183: 5589-5598
[Abstract]
[Full Text]
-
Riely, B. K., Martin, G. B.
(2001). Ancient origin of pathogen recognition specificity conferred by the tomato disease resistance gene Pto. Proc. Natl. Acad. Sci. USA
98: 2059-2064
[Abstract]
[Full Text]
-
WINSTANLEY, C., HART, C. A.
(2001). Type III secretion systems and pathogenicity islands. J Med Microbiol
50: 116-126
[Abstract]
[Full Text]
-
Mudgett, M. B., Chesnokova, O., Dahlbeck, D., Clark, E. T., Rossier, O., Bonas, U., Staskawicz, B. J.
(2000). Molecular signals required for type III secretion and translocation of the Xanthomonas campestris AvrBs2 protein to pepper plants. Proc. Natl. Acad. Sci. USA
10.1073/pnas.230450797v1
[Abstract]
[Full Text]
-
Ullrich, M. S., Schergaut, M., Boch, J., Ullrich, B.
(2000). Temperature-responsive genetic loci in the plant pathogen Pseudomonas syringae pv. glycinea. Microbiology
146: 2457-2468
[Abstract]
[Full Text]
-
Hirano, S. S., Upper, C. D.
(2000). Bacteria in the Leaf Ecosystem with Emphasis on Pseudomonas syringae---a Pathogen, Ice Nucleus, and Epiphyte. Microbiol. Mol. Biol. Rev.
64: 624-653
[Abstract]
[Full Text]
-
Collmer, A., Badel, J. L., Charkowski, A. O., Deng, W.-L., Fouts, D. E., Ramos, A. R., Rehm, A. H., Anderson, D. M., Schneewind, O., van Dijk, K., Alfano, J. R.
(2000). Pseudomonas syringae Hrp type III secretion system and effector proteins. Proc. Natl. Acad. Sci. USA
97: 8770-8777
[Abstract]
[Full Text]
-
Hendrickson, E. L., Guevera, P., Peñaloza-Vàzquez, A., Shao, J., Bender, C., Ausubel, F. M.
(2000). Virulence of the Phytopathogen Pseudomonas syringae pv. Maculicola Is rpoN Dependent. J. Bacteriol.
182: 3498-3507
[Abstract]
[Full Text]
-
Hendrickson, E. L., Guevera, P., Ausubel, F. M.
(2000). The Alternative Sigma Factor RpoN Is Required for hrp Activity in Pseudomonas syringae pv. Maculicola and Acts at the Level of hrpL Transcription. J. Bacteriol.
182: 3508-3516
[Abstract]
[Full Text]
-
Chen, L., Li, C. M., Nester, E. W.
(2000). Transferred DNA (T-DNA)-associated proteins of Agrobacterium tumefaciens are exported independently of virB. Proc. Natl. Acad. Sci. USA
10.1073/pnas.120156997v1
[Abstract]
[Full Text]
-
Alfano, J. R., Charkowski, A. O., Deng, W.-L., Badel, J. L., Petnicki-Ocwieja, T., van Dijk, K., Collmer, A.
(2000). The Pseudomonas syringae Hrp pathogenicity island has a tripartite mosaic structure composed of a cluster of type III secretion genes bounded by exchangeable effector and conserved effector loci that contribute to parasitic fitness and pathogenicity in plants. Proc. Natl. Acad. Sci. USA
97: 4856-4861
[Abstract]
[Full Text]
-
Carrington, J. C., Bisseling, T., Collmer, A., Jones, J. D.G.
(1999). Highlights from the Ninth International Congress on Molecular Plant-Microbe Interactions. Plant Cell
11: 2063-2069
[Full Text]
-
Wei, W., Plovanich-Jones, A., Deng, W.-L., Jin, Q.-L., Collmer, A., Huang, H.-C., He, S. Y.
(2000). The gene coding for the Hrp pilus structural protein is required for type III secretion of Hrp and Avr proteins in Pseudomonas syringae pv. tomato. Proc. Natl. Acad. Sci. USA
97: 2247-2252
[Abstract]
[Full Text]
-
Chen, L., Li, C. M., Nester, E. W.
(2000). Transferred DNA (T-DNA)-associated proteins of Agrobacterium tumefaciens are exported independently of virB. Proc. Natl. Acad. Sci. USA
97: 7545-7550
[Abstract]
[Full Text]
-
Mudgett, M. B., Chesnokova, O., Dahlbeck, D., Clark, E. T., Rossier, O., Bonas, U., Staskawicz, B. J.
(2000). Molecular signals required for type III secretion and translocation of the Xanthomonas campestris AvrBs2 protein to pepper plants. Proc. Natl. Acad. Sci. USA
97: 13324-13329
[Abstract]
[Full Text]