RsmA regulates biofilm formation in Xanthomonas campestris through a regulatory network involving cyclic di-GMP and the Clp transcription factor

PLoS One. 2012;7(12):e52646. doi: 10.1371/journal.pone.0052646. Epub 2012 Dec 21.

Abstract

Biofilm formation and dispersal in the black rot pathogen Xanthomonas campestris pathovar campestris (Xcc) is influenced by a number of factors. The extracellular mannanase ManA has been implicated in biofilm dispersal whereas biofilm formation requires a putative glycosyl transferase encoded by the xag gene cluster. Previously we demonstrated that the post-transcriptional regulator RsmA exerts a negative regulatory influence on biofilm formation in Xcc. Here we address the mechanisms by which RsmA exerts this action. We show that RsmA binds to the transcripts of three genes encoding GGDEF domain diguanylate cyclases to influence their expression. Accordingly, mutation of rsmA leads to an increase in cellular levels of cyclic di-GMP. This effect is associated with a down-regulation of transcription of manA, but an upregulation of xag gene transcription. Mutation of clp, which encodes a cyclic di-GMP-responsive transcriptional regulator of the CRP-FNR family, has similar divergent effects on the expression of manA and xag. Nevertheless Clp binding to manA and xag promoters is inhibited by cyclic di-GMP. The data support the contention that, in common with other CRP-FNR family members, Clp can act as both an activator and repressor of transcription of different genes to influence biofilm formation as a response to cyclic di-GMP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism*
  • Biofilms*
  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / metabolism
  • Gene Expression Regulation, Bacterial
  • Models, Biological
  • Mutation
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Interaction Domains and Motifs / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • Xanthomonas campestris / genetics
  • Xanthomonas campestris / metabolism*

Substances

  • Bacterial Proteins
  • Clp protein, Xanthomonas campestris
  • RNA, Messenger
  • Transcription Factors
  • bis(3',5')-cyclic diguanylic acid
  • Cyclic GMP

Grants and funding

The work of the authors has been supported in part by grants awarded by the Science Foundation Ireland (SFI 07/IN.1/B955 to J.M.D. and SFI 09/SIRG/B1654 to R.P.R.), Royal Irish Academy fellowship (to R.R.P) and an International Research Partnership supported by Science Foundation Ireland (07/IN.1/B955/IRPs) and the National Science Foundation of China (30810103912). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.