A complete twelve-gene deletion null mutant reveals that cyclic di-GMP is a global regulator of phase-transition and host colonization in Erwinia amylovora

PLoS Pathog. 2022 Aug 1;18(8):e1010737. doi: 10.1371/journal.ppat.1010737. eCollection 2022 Aug.

Abstract

Cyclic-di-GMP (c-di-GMP) is an essential bacterial second messenger that regulates biofilm formation and pathogenicity. To study the global regulatory effect of individual components of the c-di-GMP metabolic system, we deleted all 12 diguanylate cyclase (dgc) and phosphodiesterase (pde)-encoding genes in E. amylovora Ea1189 (Ea1189Δ12). Ea1189Δ12 was impaired in surface attachment due to a transcriptional dysregulation of the type IV pilus and the flagellar filament. A transcriptomic analysis of surface-exposed WT Ea1189 and Ea1189Δ12 cells indicated that genes involved in metabolism, appendage generation and global transcriptional/post-transcriptional regulation were differentially regulated in Ea1189Δ12. Biofilm formation was regulated by all 5 Dgcs, whereas type III secretion and disease development were differentially regulated by specific Dgcs. A comparative transcriptomic analysis of Ea1189Δ8 (lacks all five enzymatically active dgc and 3 pde genes) against Ea1189Δ8 expressing specific dgcs, revealed the presence of a dual modality of spatial and global regulatory frameworks in the c-di-GMP signaling network.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms
  • Cyclic GMP / analogs & derivatives
  • Cyclic GMP / genetics
  • Cyclic GMP / metabolism
  • Erwinia amylovora* / genetics
  • Erwinia amylovora* / metabolism
  • Escherichia coli Proteins* / metabolism
  • Gene Deletion
  • Gene Expression Regulation, Bacterial
  • Phosphoric Diester Hydrolases / metabolism
  • Phosphorus-Oxygen Lyases / genetics
  • Phosphorus-Oxygen Lyases / metabolism

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • bis(3',5')-cyclic diguanylic acid
  • Phosphoric Diester Hydrolases
  • Phosphorus-Oxygen Lyases
  • Cyclic GMP

Grants and funding

This project was supported by funds from the Agriculture and Food Research Initiative Competitive Grants Program Grants no. 2015-67013-23068 (GWS) and 2020-51181-32158 (GWS) from the USDA National Institute of Food and Agriculture, and by Michigan State University AgBioResearch (GWS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.