Mutational analysis of Xanthomonas harpin HpaG identifies a key functional region that elicits the hypersensitive response in nonhost plants

Journal of Bacteriology
Jung-Gun KimIngyu Hwang

Abstract

HpaG is a type III-secreted elicitor protein of Xanthomonas axonopodis pv. glycines. We have determined the critical amino acid residues important for hypersensitive response (HR) elicitation by random and site-directed mutagenesis of HpaG and its homolog XopA. A plasmid clone carrying hpaG was mutagenized by site-directed mutagenesis, hydroxylamine mutagenesis, and error-prone PCR. A total of 52 mutants were obtained, including 51 single missense mutants and 1 double missense mutant. The HR elicitation activity was abolished in the two missense mutants [HpaG(L50P) and HpaG(L43P/L50P)]. Seven single missense mutants showed reduced activity, and the HR elicitation activity of the rest of the mutants was similar to that of wild-type HpaG. Mutational and deletion analyses narrowed the region essential for elicitor activity to the 23-amino-acid peptide (H2N-NQGISEKQLDQLLTQLIMALLQQ-COOH). A synthetic peptide of this sequence possessed HR elicitor activity at the same concentration as the HpaG protein. This region has 78 and 74% homology with 23- and 27-amino-acid regions of the HrpW harpin domains, respectively, from Pseudomonas and Erwinia spp. The secondary structure of the peptide is predicted to be an alpha-helix, as is the HrpW...Continue Reading

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Citations

Apr 6, 2005·FEMS Microbiology Reviews·Gail M PrestonIsabelle Caldelari
Jun 12, 2013·Molecular Plant-microbe Interactions : MPMI·Min-Seon ChoiChang-Sik Oh
Mar 4, 2006·Journal of Bacteriology·Mee-Ngan YapAmy O Charkowski
Sep 10, 2013·Journal of Experimental Botany·Markus Albert
Oct 26, 2013·Molecular Plant-microbe Interactions : MPMI·E VergneE Chevreau
Sep 4, 2014·T︠S︡itologii︠a︡ i genetika·S N Shamraĭ
Apr 1, 2008·Cellular Microbiology·Elisabeth Ashman Epstein, Matthew R Chapman
Jul 14, 2012·Molecular Plant Pathology·Germán G SgroJorgelina Ottado
May 8, 2010·Molecular Plant Pathology·Jian-Gang LiJian-Hua Guo
Aug 20, 2014·Frontiers in Microbiology·Adnan K Syed, Blaise R Boles
Feb 6, 2013·International Journal of Molecular Sciences·Inés Ponce de León, Marcos Montesano
Feb 23, 2007·The Journal of Biological Chemistry·Jonghee OhIngyu Hwang
May 3, 2019·Proteomics·Fabiano T P K TávoraAngela Mehta
Oct 4, 2020·International Journal of Molecular Sciences·Anastasiia O KosolapovaAnton A Nizhnikov
Aug 19, 2020·Cell Host & Microbe·Neeraj K LalSavithramma P Dinesh-Kumar

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