DOI: 10.1111/j.1364-3703.2010.00655.x. Functional mapping of harpin HrpZ of<i>Pseudomonas syringae</i>reveals the sites responsible for protein oligomerization, lipid interactions and plant defence induction

Functional mapping of harpin HrpZ ofPseudomonas syringaereveals the sites responsible for protein oligomerization, lipid interactions and plant defence induction

10.1111/j.1364-3703.2010.00655.x

Crossref journal-article

Wiley

Molecular Plant Pathology (311)

Abstract

SUMMARYHarpin HrpZ is one of the most abundant proteins secreted through the pathogenesis‐associated type III secretion system of the plant pathogenPseudomonas syringae. HrpZ shows membrane‐binding and pore‐forming activitiesin vitro, suggesting that it could be targeted to the host cell plasma membrane. We studied the native molecular forms of HrpZ and found that it forms dimers and higher order oligomers. Lipid binding by HrpZ was tested with 15 different membrane lipids, with HrpZ interacting only with phosphatidic acid. Pore formation by HrpZ in artificial lipid vesicles was found to be dependent on the presence of phosphatidic acid. In addition, HrpZ was able to form pores in vesicles prepared fromArabidopsis thalianaplasma membrane, providing evidence for the suggested target of HrpZ in the host. To map the functions associated with HrpZ, we constructed a comprehensive series of deletions in thehrpZgene derived fromP. syringaepv.phaseolicola, and studied the mutant proteins. We found that oligomerization is mainly mediated by a region near the C‐terminus of the protein, and that the same region is also essential for membrane pore formation. Phosphatidic acid binding seems to be mediated by two regions separate in the primary structure. Tobacco, a nonhost plant, recognizes, as a defence elicitor, a 24‐amino‐acid HrpZ fragment which resides in the region indispensable for the oligomerization and pore formation functions of HrpZ.

Bibliography

HAAPALAINEN, M., ENGELHARDT, S., KÃFNER, I., LI, C., NÃRNBERGER, T., LEE, J., ROMANTSCHUK, M., & TAIRA, S. (2010). Functional mapping of harpin HrpZ ofPseudomonas syringaereveals the sites responsible for protein oligomerization, lipid interactions and plant defence induction. Molecular Plant Pathology, 12(2), 151â166. Portico.

Authors 8

MINNA HAAPALAINEN (first)
STEFAN ENGELHARDT (additional)
ISABELL KÜFNER (additional)
CHUN‐MEI LI (additional)
THORSTEN NÜRNBERGER (additional)
JUSTIN LEE (additional)
MARTIN ROMANTSCHUK (additional)
SUVI TAIRA (additional)

References 42 Referenced 52

10.1046/j.1365-2958.1996.415946.x
10.1016/j.bbalip.2009.04.006
10.1083/jcb.147.3.683
10.1111/j.1364-3703.2008.00471.x
10.1046/j.1364-3703.2002.00139.x
10.1094/MPMI.2001.14.3.394
10.1128/JB.184.9.2389-2398.2002
10.1128/JB.180.19.5211-5217.1998
10.1006/pmpp.1997.0120
10.1006/prep.1994.1020
10.1111/j.1365-313X.2008.03723.x
10.1146/annurev.micro.60.080805.142251
10.1094/MPMI-22-9-1069
10.1094/MPMI-22-3-0282
10.1002/j.1460-2075.1996.tb00968.x
10.1016/0092-8674(93)90354-S
10.1126/science.1066397
10.1128/JB.186.18.6239-6247.2004
10.1074/jbc.M609737200
10.1128/JB.01146-07
10.1105/tpc.13.5.1079
10.1073/pnas.011265298
10.1093/emboj/21.8.1909
10.1094/MPMI-18-0060
10.1134/S1021443706010055
10.1093/jxb/erm239
10.1073/pnas.0902362106
10.1128/JB.187.2.649-663.2005
10.1094/MPMI-8-0717
10.1111/j.1365-2958.2004.04910.x
10.1042/0264-6021:3420119
10.1073/pnas.94.7.3459
10.1016/S1016-8478(23)17547-9 / Mol. Cells / Genetic organization of the hrp genes cluster in Erwinia pyrifoliae and characterization of HR active domains in HrpNEp protein by mutational analysis by Shrestha R. (2008)
10.1094/MPMI-21-11-1387
10.1016/j.bbalip.2006.03.006
10.1007/PL00012997
10.1094/MPMI.2000.13.12.1366
10.1016/j.bbapap.2009.07.023
10.1016/j.tplants.2005.06.002
10.1094/MPMI.2004.17.11.1250
10.1104/pp.109.1.15
10.1007/s11033-006-9034-6

Dates

Type	When
Created	15 years ago (Aug. 17, 2010, 3:01 a.m.)
Deposited	1 year, 5 months ago (March 29, 2024, 1:13 p.m.)
Indexed	3 days ago (Aug. 28, 2025, 8:03 a.m.)
Issued	15 years ago (Aug. 17, 2010)
Published	15 years ago (Aug. 17, 2010)
Published Online	15 years ago (Aug. 17, 2010)
Published Print	14 years, 6 months ago (Feb. 1, 2011)

Funders 0

None

BibTeX

@article{HAAPALAINEN_2010, title={Functional mapping of harpin HrpZ ofPseudomonas syringaereveals the sites responsible for protein oligomerization, lipid interactions and plant defence induction}, volume={12}, ISSN={1364-3703}, url={http://dx.doi.org/10.1111/j.1364-3703.2010.00655.x}, DOI={10.1111/j.1364-3703.2010.00655.x}, number={2}, journal={Molecular Plant Pathology}, publisher={Wiley}, author={HAAPALAINEN, MINNA and ENGELHARDT, STEFAN and KÜFNER, ISABELL and LI, CHUN‐MEI and NÜRNBERGER, THORSTEN and LEE, JUSTIN and ROMANTSCHUK, MARTIN and TAIRA, SUVI}, year={2010}, month=aug, pages={151–166} }

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