Molecular biology of Clostridium difficile

Principal investigator(s)

Dr. J. (Jeroen) CorverDr. W.K. (Wiep Klaas) Smits

Description of the research

Pathogenic Clostridium difficile strains produce several known virulence factors of which the most well-known are the toxins TcdA and TcdB, and the binary toxin. Toxin expression is positively regulated by the sigma factor TcdR and negatively by the anti-sigma factor TcdC. We investigate the virulence/pathogenesis of C. difficile by characterizing the biochemical properties of TcdC such as dimerization, membrane association, and binding to TcdR. In addition, we aim to identify novel virulence factors through genomic and proteomic approaches. Using these approaches we are also searching for molecular markers that are representative for hypervirulent types of C. difficile. By investigating the signaling pathways and gene networks that regulate these factors and the effects of these factors on the host environment using genetics, molecular biology, cell biology and biochemistry we aim to identify potential targets for the development of novel antibiotics.

Research objectives

  • Study the regulation of toxin expression in C. difficile
  • Identify novel virulence factors of C. difficile andmolecular markers for C. difficile hypervirulence
  • Determine genome wide binding profiles of transcription factors involved in C. difficile virulence and pathogenesis
  • Characterize the DNA replication machinery of C. difficile
  • Investigate the effect of extracellular C. difficile factors on the host environment

Key publications

  • Diet driven selection of epidemic, hypervirulent ribotypes of an enteric pathogen. Collins J, Robinson C, Danhof H,  Knetsch CW, van Leeuwen HC, Lawley TD, Auchtung JM and Britton RA. Nature. 2017. In press.
  • A helicase-containing module defines a family of pCD630-like plasmids in Clostridium difficile. Wiep Klaas Smits,  J. Scott Weese, Adam P. Roberts, Céline Harmanus, Bastian Hornung. Anaerobe. 2017. In press. doi:10.1016/j.anaerobe.2017.12.005
  • Covalent attachment and Pro-Pro endopeptidase (PPEP-1)-mediated release of Clostridium difficile cell surface proteins involved in adhesion. Corver J, Cordo' V, van Leeuwen HC, Klychnikov OI, Hensbergen PJ. Mol Microbiol. 2017 Sep;105(5):663-673 doi: 10.1111/mmi.13736.
  • The Signal Sequence of the Abundant Extracellular Metalloprotease PPEP-1 Can Be Used to Secrete Synthetic Reporter Proteins in Clostridium difficile. Oliveira Paiva AM, Friggen AH, Hossein-Javaheri S, Smits WK. ACS Synth Biol. 2016 Dec 16;5(12):1376-1382
  • Primase is required for helicase activity and helicase alters the specificity of primase in the enteropathogen Clostridium difficile. Open Biol. 2016; 6:160272 doi: 10.1098/rsob.160272.
  • A Novel Fic (Filamentation Induced by cAMP) Protein from Clostridium difficile Reveals an Inhibitory Motif-independent Adenylylation/AMPylation Mechanism. Dedic E, Alsarraf H, Welner DH, Østergaard O, Klychnikov OI, Hensbergen PJ, Corver J, van Leeuwen HC, Jørgensen R. J Biol Chem. 2016 Jun 17;291(25):13286-300 doi: 10.1074/jbc.M115.705491.
  • Complete genome sequence of the Clostridium difficile laboratory strain 630Δerm reveals differences from strain 630, including translocation of the mobile element CTn5. van Eijk E, Anvar SY, Browne HP, Leung WY, Frank J, Schmitz AM, Roberts AP, Smits WK. BMC Genomics. 2015 Jan 31;16:31. doi: 10.1186/s12864-015-1252-7.
  • The HtrA-like protease CD3284 modulates virulence of Clostridium difficile. Bakker D, Buckley AM, de Jong A, van Winden VJ, Verhoeks JP, Kuipers OP, Douce GR, Kuijper EJ, Smits WK, Corver J. Infect Immun. 2014 Oct;82(10):4222-32 doi: 10.1128/IAI.02336-14.
  • C. difficile 630Δerm Spo0A regulates sporulation, but does not contribute to toxin production, by direct high-affinity binding to target DNA. Rosenbusch KE, Bakker D, Kuijper EJ, Smits WK. PLoS One. 2012; 7:e48608 doi: 10.1371/journal.pone.0048608.
  • TcdC does not significantly repress toxin expression in Clostridium difficile 630ΔErm. Bakker D, Smits WK, Kuijper EJ, Corver J. PLoS One. 2012; 7:e43247 doi: 10.1371/journal.pone.0043247.

Research group

  • Ilse Boekhoud, MSc - PhD student
  • Jeroen Corver, PhD - PI, Molecular Biologist
  • Erika van Eijk, MD - PhD student
  • Annemieke Friggen, BSc - Technician
  • Ed Kuijper, PhD - Medical Microbiologist
  • Ana Paiva, MSc - PhD student
  • Wiep Klaas Smits, PhD - Assistant professor, Molecular Biologist

Collaborators

  • Paul Hensbergen (LUMC)
  • Adam Roberts (Liverpool School of Tropical Medicine)
  • Brian Mark (University of Manitoba)
  • Oleg Klychnikov (KU Leuven)
  • Gillian Douce (Glasgow)
  • Robert Fagan (University of Sheffield)