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  • 1
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 25, No. 2 ( 2012-02), p. 231-240
    Abstract: The type III effector HsvG of the gall-forming Pantoea agglomerans pv. gypsophilae is a DNA-binding protein that is imported to the host nucleus and involved in host specificity. The DNA-binding region of HsvG was delineated to 266 amino acids located within a secondary structure region near the N-terminus of the protein but did not display any homology to canonical DNA-binding motifs. A binding site selection procedure was used to isolate a target gene of HsvG, named HSVGT, in Gypsophila paniculata. HSVGT is a predicted acidic protein of the DnaJ family with 244 amino acids. It harbors characteristic conserved motifs of a eukaryotic transcription factor, including a bipartite nuclear localization signal, zinc finger, and leucine zipper DNA-binding motifs. Quantitative real-time polymerase chain reaction analysis demonstrated that HSVGT transcription is specifically induced in planta within 2 h after inoculation with the wild-type P. agglomerans pv. gypsophilae compared with the hsvG mutant. Induction of HSVGT reached a peak of sixfold at 4 h after inoculation and progressively declined thereafter. Gel-shift assay demonstrated that HsvG binds to the HSVGT promoter, indicating that HSVGT is a direct target of HsvG. Our results support the hypothesis that HsvG functions as a transcription factor in gypsophila.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2012
    detail.hit.zdb_id: 2037108-1
    SSG: 12
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  • 2
    Online Resource
    Online Resource
    Scientific Societies ; 2018
    In:  Molecular Plant-Microbe Interactions® Vol. 31, No. 2 ( 2018-02), p. 233-239
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 31, No. 2 ( 2018-02), p. 233-239
    Abstract: Salmonella enterica serovar Typhimurium, a human enteric pathogen, has the ability to multiply and survive endophytically in plants. Genes encoding the type III secretion system (T3SS) or its effectors (T3Es) may contribute to its colonization. Two reporter plasmids for T3E translocation into plant cells that are based on hypersensitive response domains of avirulence proteins from the Pantoea agglomerans–beet and Xanthomonas euvesicatoria–pepper pathosystems were employed in this study to investigate the role of T3Es in the interaction of Salmonella ser. Typhimurium 14028 with plants. The T3Es of Salmonella ser. Typhimurium, SipB and SifA, which are translocated into animal cells, could not be delivered by Salmonella ser. Typhimurium into cells of beet roots or pepper leaves. In contrast, these effectors were translocated into plant cells by the phytopathogenic bacteria P. agglomerans pv. betae, Erwinia amylovora, and X. euvesicatoria. Similarly, HsvG, a T3E of P. agglomerans pv. gypsophilae, and XopAU of X. euvesicatoria could be translocated into beet roots and pepper leaves, respectively, by the plant pathogens but not by Salmonella ser. Typhimurium. Mutations in Salmonella ser. Typhimurium T3SS genes invA, ssaV, sipB, or sifA, did not affect its endophytic colonization of lettuce leaves, supporting the notion that S. enterica cannot translocate T3Es into plant cells.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2018
    detail.hit.zdb_id: 2037108-1
    SSG: 12
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  • 3
    In: Molecular Microbiology, Wiley, Vol. 61, No. 5 ( 2006-09), p. 1118-1131
    Abstract: Pantoea agglomerans pv. gypsophilae ( Pag ) elicits galls on gypsophila and a hypersensitive response on beet, whereas P. agglomerans pv. betae ( Pab ) induces galls on both beet and gypsophila. The pathogenicity of both pathovars is dependent on the presence of a plasmid harbouring type III secretion system (TTSS) components and effectors. The HsvG TTSS effectors of Pag (HsvG‐ Pag ) and Pab (HsvG‐ Pab ) determine the host specificity of both pathovars on gypsophila. Here we describe a novel HsvG homologue, HsvB, which determines the host specificity of Pag and Pab on beet. HsvG requires two direct amino acid repeats for pathogenicity on gypsophila, whereas one repeat in HsvB is sufficient for pathogenicity on beet. Exchanging repeats between HsvG‐ Pag and HsvB‐ Pab resulted in a switch of host specificities. Transient expression of GFP–HsvG or GFP–HsvB fusions in gypsophila, beet or melon leaves showed that HsvG and HsvB were localized to the nuclei of host and non‐host plants. A yeast one‐hybrid assay revealed that a single repeat of HsvG or HsvB was sufficient to activate transcription. By employing random binding‐site selection and gel‐shift assay HsvG was demonstrated to be a double‐stranded DNA‐binding protein with an ACACC/aAA consensus binding site. These results suggest that HsvG and HsvB are host‐specificity determinants and bear the potential to affect the host transcriptional machinery.
    Type of Medium: Online Resource
    ISSN: 0950-382X , 1365-2958
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2006
    detail.hit.zdb_id: 1501537-3
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  • 4
    Online Resource
    Online Resource
    Scientific Societies ; 2005
    In:  Molecular Plant-Microbe Interactions® Vol. 18, No. 7 ( 2005-07), p. 634-643
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 18, No. 7 ( 2005-07), p. 634-643
    Abstract: HrpL, an alternative σ factor, activates the transcription of the Hrp regulon by its binding to a common “hrp box” promoter. Based on computational techniques, the hrp box previously was defined as a consensus bipartite cis element, 5′-GGAACC-N 15–16 -CCACNNA-3′. The present report combines a quantitative in vivo assay for measuring Hrp promoter activity with site-specific mutagenesis to analyze the effect of consensus and nonconsensus nucleotides on promoter activity. The analysis was carried out with Hop effectors of the tumorigenic bacterium Pantoea agglomerans pv. gypsophilae, in which HrpL is indispensable for gall formation. Mutational analysis indicates that the hrp box consensus can be divided into crucial and noncrucial nucleotides. The first 5 nucleotides (nt) of the -35 consensus motif (GGAAC) and the 3 nt of the -10 motif (ACNNA) are crucial, whereas other consensus and adjacent nonconsensus nucleotides exert a significant effect on the promoter's strength. With spacing of 13 or 17 nt between the two motifs, significant activity was still retained. Gel shift assays indicated that deletion of GG from the -35 consensus motif eliminated HrpL binding, whereas mutations in the -10 consensus motif or modification of the spacing, which eliminates promoter activity, did not elicit any effect. The degeneracy in Hrp promoters of four hrp and type III effector genes of P. agglomerans pv. gypsophilae indicated significant differences in promoter activity, whereas increasing the promoter strength of the Hop effector, HsvG, resulted in overexpression of gall formation.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2005
    detail.hit.zdb_id: 2037108-1
    SSG: 12
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  • 5
    In: Molecular Plant Pathology, Wiley, Vol. 19, No. 2 ( 2018-02), p. 381-392
    Abstract: Pantoea agglomerans , a widespread epiphytic bacterium, has evolved into a hypersensitive response and pathogenicity ( hrp )‐dependent and host‐specific gall‐forming pathogen by the acquisition of a pathogenicity plasmid containing a type III secretion system (T3SS) and its effectors (T3Es). Pantoea agglomerans pv. betae ( Pab ) elicits galls on beet ( Beta vulgaris ) and gypsophila ( Gypsophila paniculata ), whereas P. agglomerans pv. gypsophilae ( Pag ) incites galls on gypsophila and a hypersensitive response (HR) on beet. Draft genome sequences were generated and employed in combination with a machine‐learning approach and a translocation assay into beet roots to identify the pools of T3Es in the two pathovars. The genomes of the sequenced Pab 4188 and Pag 824‐1 strains have a similar size (∼5 MB) and GC content (∼55%). Mutational analysis revealed that, in Pab 4188, eight T3Es (HsvB, HsvG, PseB, DspA/E, HopAY1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on beet and gypsophila. In Pag 824‐1, nine T3Es (HsvG, HsvB, PthG, DspA/E, HopAY1, HopD1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on gypsophila, whereas the PthG effector triggers HR on beet. HsvB, HsvG, PthG and PseB appear to endow pathovar specificities to Pab and Pag , and no homologous T3Es were identified for these proteins in other phytopathogenic bacteria. Conversely, the remaining T3Es contribute to the virulence of both pathovars, and homologous T3Es were found in other phytopathogenic bacteria. Remarkably, HsvG and HsvB, which act as host‐specific transcription factors, displayed the largest contribution to disease development.
    Type of Medium: Online Resource
    ISSN: 1464-6722 , 1364-3703
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2018
    detail.hit.zdb_id: 2020755-4
    SSG: 12
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  • 6
    In: Molecular Plant Pathology, Wiley, Vol. 20, No. 11 ( 2019-11), p. 1582-1587
    Abstract: Pantoea agglomerans ( Pa ), a widespread commensal bacterium, has evolved into a host‐specific gall‐forming pathogen on gypsophila and beet by acquiring a plasmid harbouring a type III secretion system (T3SS) and effectors (T3Es). Pantoea agglomerans pv. gypsophilae ( Pag ) elicits galls on gypsophila and a hypersensitive response on beet, whereas P. agglomerans pv. betae ( Pab ) elicits galls on beet and gypsophila. HsvG and HsvB are two paralogous T3Es present in both pathovars and act as host‐specific transcription activators on gypsophila and beet, respectively. PthG and PseB are major T3Es that contribute to gall development of Pag and Pab , respectively. To establish the minimal combinations of T3Es that are sufficient to elicit gall symptoms, strains of the nonpathogenic bacteria Pseudomonas fluorescens 55, Pa 3‐1, Pa 98 and Escherichia coli , transformed with pHIR11 harbouring a T3SS, and the phytopathogenic bacteria Erwinia amylovora , Dickeya solani and Xanthomonas campestris pv. campestris were transformed with the T3Es hsvG , hsvB , pthG and pseB , either individually or in pairs, and used to infect gypsophila and beet. Strikingly, all the tested nonpathogenic and phytopathogenic bacterial strains harbouring hsvG and pthG incited galls on gypsophila, whereas strains harbouring hsvB and pseB , with the exception of E. coli , incited galls on beet.
    Type of Medium: Online Resource
    ISSN: 1464-6722 , 1364-3703
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2019
    detail.hit.zdb_id: 2020755-4
    SSG: 12
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