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Characterization of hns genes from Erwinia amylovora

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Abstract

The small basic histone-like protein H-NS is known for bacteria to attenuate virulence of several animal pathogens. An hns homologue from E. amylovora was identified by complementing an E. coli hns-mutant strain with a cosmid library from E. amylovora. A 1.6 kb EcoRI-fragment complemented the mucoid phenotype and repressed the ß-glucosidase activity of E. coli PD32. The open reading frame encoding an H-NS-like protein of 134 amino acid was later shown to be located on plasmid pEA29 (McGhee and Jones 2000). A chromosomal hns gene was amplified with PCR consensus primers and localized near galU of E. amylovora. E. amylovora mutants were created by insertion of a resistance cassette, and the intact gene was inserted into a high copy number plasmid for constitutive expression. Purified chromosomal H-NS protein preferentially bound to a DNA fragment from the lsc region and bending was predicted for an adjacent fragment with the rlsB-promoter. Levan production was significantly increased by hns mutations. Synthesis of the capsular exopolysaccharide amylovoran and of levan were reduced, when hns from the E. amylovora plasmid was overexpressed. A mutation in chromosomal hns of E. amylovora increased amylovoran synthesis, and both mutations retarded symptom formation on immature pears.

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Acknowledgements

We thank S. Jock for a series of experiments to cure cells from pEA29 and B. Schneider for comments on the manuscript.

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Authors and Affiliations

  1. Max-Planck-Institut für Zellbiologie, Ladenburg, Germany

    M. Hildebrand, P. Aldridge & K. Geider

  2. BBA Dossenheim, Schwabenheimer Strasse 101, 69221, Dossenheim, Germany

    K. Geider

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  1. M. Hildebrand
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  2. P. Aldridge
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  3. K. Geider
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Correspondence to K. Geider.

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Communicated by A. Kondorosi

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Hildebrand, M., Aldridge, P. & Geider, K. Characterization of hns genes from Erwinia amylovora . Mol Genet Genomics 275, 310–319 (2006). https://doi.org/10.1007/s00438-005-0085-5

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