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Biofilm formation in Escherichia coli cra mutants is impaired due to down-regulation of curli biosynthesis

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Abstract

Cra is a pleiotropic regulatory protein that controls carbon and energy flux in enteric bacteria. Recent studies have shown that Cra also regulates other cell processes and influences biofilm formation. The purpose of the present study was to investigate the role of Cra in biofilm formation in Escherichia coli. Congo red-binding studies suggested that curli biosynthesis is impaired in cra mutants. Microarray analysis of wild-type and mutant E. coli cultivated in conditions promoting biofilm formation revealed that the curli biosynthesis genes, csgBAC and csgDEFG, are poorly expressed in the mutant, suggesting that transcription of genes required for curli production is regulated by Cra. Four putative Cra-binding sites were identified in the curli intergenic region, which were experimentally validated by performing electromobility shift assays. Site-directed mutagenesis of three Cra-binding sites in the promoter region of the csgDEFG operon suggests that Cra activates transcription of this operon upon binding to operator regions both downstream and upstream of the transcription start site. Based on the Cra-binding sites identified in this and other studies, the Cra consensus sequence is refined.

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Acknowledgments

We thank Prof. P. Jayadeva Bhat for helpful discussions and insights. S.M.S.R. is a recipient of a research fellowship from the Indian Council of Medical Research.

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Authors and Affiliations

  1. Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai, Maharashtra, 400076, India

    Shamlan M. S. Reshamwala

  2. Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai, Maharashtra, 400076, India

    Santosh B. Noronha

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  1. Shamlan M. S. Reshamwala
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  2. Santosh B. Noronha
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Correspondence to Santosh B. Noronha.

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Communicated by Jorge Membrillo-Hernández.

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Reshamwala, S.M.S., Noronha, S.B. Biofilm formation in Escherichia coli cra mutants is impaired due to down-regulation of curli biosynthesis. Arch Microbiol 193, 711–722 (2011). https://doi.org/10.1007/s00203-011-0708-7

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