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Identification of the flagellar chaperone FlgN in the phytopathogen Xanthomonas axonopodis pathovar citri by its interaction with hook-associated FlgK

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Abstract

Genome annotation of the plant pathogen Xanthomonas axonopodis pv. citri (Xac), identified flagellar genes in a 15.7 kb gene cluster. However, FlgN, a secretion chaperone for hook-associated proteins FlgK and FlgL, was not identified. We performed extensive screening of the X. axonopodis pv. citri genome with the yeast two-hybrid system to identify a protein with the characteristics of the flagellar chaperone FlgN. We found a candidate (XAC1990) encoded by an operon for components of the flagellum apparatus that interacted with FlgK. In order to further support this finding, Xac FlgK and XAC1990 were cloned, expressed, and purified. The recombinant proteins were characterized by spectroscopic methods and their interaction in vitro confirmed by pull-down assays. We, therefore, conclude that XAC1990 and its homologs in other Xanthomonas species are, in fact, FlgN proteins. These observations extend the sequence diversity covered by this family of proteins.

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Abbreviations

Xac :

Xanthomonas axonopodis pathovar citri

3AT:

3-Aminotriazole

MM:

Molecular mass

A:

Absorbance

CD:

Circular dichroism

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Acknowledgments

We thank Nicola Conran for a critical reading of this manuscript. We are grateful for the excellent technical assistance of Silvia da Silva, Veruska Soares, and Ivanilce Guimarães. We thank Professor Stanley Fields for kindly providing the pOBD and pOAD plasmids and Professor Phillip James for kindly providing PJ694-a yeast cells. This study was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). LK, MCA, TMS, LT and CD thank FAPESP for fellowship.

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Authors and Affiliations

  1. Laboratório Nacional de Luz Síncrotron, P.O. Box 6192, CEP 13084-971, Campinas, SP, Brazil

    Letícia Khater, Paula F. L. Borin, Túlio M. Santos, Ljubica Tasic & Carlos H. I. Ramos

  2. Universidade Estadual de Campinas, CEP 13083-970, Campinas, SP, Brazil

    Letícia Khater, Paula F. L. Borin, Ljubica Tasic & Carlos H. I. Ramos

  3. Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05508-000, São Paulo, SP, Brazil

    Marcos C. Alegria, Cássia Docena & Chuck S. Farah

  4. UNICAMP Instituto de Quimica, PO Box 6154, 13084-862, Campinas, SP, Brazil

    Carlos H. I. Ramos

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  1. Letícia Khater
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  2. Marcos C. Alegria
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  3. Paula F. L. Borin
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  4. Túlio M. Santos
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  6. Ljubica Tasic
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  7. Chuck S. Farah
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  8. Carlos H. I. Ramos
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Correspondence to Carlos H. I. Ramos.

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Communicated by Jan Tommassen.

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Khater, L., Alegria, M.C., Borin, P.F.L. et al. Identification of the flagellar chaperone FlgN in the phytopathogen Xanthomonas axonopodis pathovar citri by its interaction with hook-associated FlgK. Arch Microbiol 188, 243–250 (2007). https://doi.org/10.1007/s00203-007-0240-y

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  • DOI: https://doi.org/10.1007/s00203-007-0240-y

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