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Role of σ54 in the regulation of genes involved in type I and type IV pili biogenesis in Xylella fastidiosa

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Abstract

The phytopathogen Xylella fastidiosa produces long type IV pili and short type I pili involved in motility and adhesion. In this work, we have investigated the role of sigma factor σ54 (RpoN) in the regulation of fimbrial biogenesis in X. fastidiosa. An rpoN null mutant was constructed from the non-pathogenic citrus strain J1a12, and microarray analyses of global gene expression comparing the wild type and rpoN mutant strains showed few genes exhibiting differential expression. In particular, gene pilA1 (XF2542), which encodes the structural pilin protein of type IV pili, showed decreased expression in the rpoN mutant, whereas two-fold higher expression of an operon encoding proteins of type I pili was detected, as confirmed by quantitative RT-PCR (qRT-PCR) analysis. The transcriptional start site of pilA1 was determined by primer extension, downstream of a σ54-dependent promoter. Microarray and qRT-PCR data demonstrated that expression of only one of the five pilA paralogues, pilA1, was significantly reduced in the rpoN mutant. The rpoN mutant made more biofilm than the wild type strain and presented a cell-cell aggregative phenotype. These results indicate that σ54 differentially regulates genes involved in type IV and type I fimbrial biogenesis, and is involved in biofilm formation in X. fastidiosa.

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Acknowledgments

This work was supported by a grant from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). During the course of this work, J. F. S. N. and T. K. were supported by predoctoral fellowships from FAPESP. M. V. M. and S. L. G. are partly supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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  1. Tie Koide

    Present address: Institute for Systems Biology, 1441 North 34th Street, Seattle, WA, 98103-8904, USA

Authors and Affiliations

  1. Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1374, 05508-000, São Paulo, SP, Brazil

    José F. da Silva Neto & Marilis V. Marques

  2. Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes 748, 05508-000, São Paulo, SP, Brazil

    Tie Koide & Suely L. Gomes

  3. Laboratório de Bacteriologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900, São Paulo, SP, Brazil

    Cecília M. Abe

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  1. José F. da Silva Neto
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Correspondence to Marilis V. Marques.

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Communicated by Jack Meeks.

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da Silva Neto, J.F., Koide, T., Abe, C.M. et al. Role of σ54 in the regulation of genes involved in type I and type IV pili biogenesis in Xylella fastidiosa . Arch Microbiol 189, 249–261 (2008). https://doi.org/10.1007/s00203-007-0314-x

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