title: Type IV pili promote early biofilm formation by Clostridium difficile.
abstract: Increasing morbidity and mortality from Clostridium difficile infection (CDI) present an enormous challenge to healthcare systems. Clostridium difficile express type IV pili (T4P), but their function remains unclear. Many chronic and recurrent bacterial infections result from biofilms, surface-associated bacterial communities embedded in an extracellular matrix. CDI may be biofilm mediated; T4P are important for biofilm formation in a number of organisms. We evaluate the role of T4P in C. difficile biofilm formation using RNA sequencing, mutagenesis and complementation of the gene encoding the major pilin pilA1, and microscopy. RNA sequencing demonstrates that, in comparison to other growth phenotypes, C. difficile growing in a biofilm has a distinct RNA expression profile, with significant differences in T4P gene expression. Microscopy of T4P-expressing and T4P-deficient strains suggests that T4P play an important role in early biofilm formation. A non-piliated pilA1 mutant forms an initial biofilm of significantly reduced mass and thickness in comparison to the wild type. Complementation of the pilA1 mutant strain leads to formation of a biofilm which resembles the wild-type biofilm. These findings suggest that T4P play an important role in early biofilm formation. Novel strategies for confronting biofilm infections are emerging; our data suggest that similar strategies should be investigated in CDI.
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Grace A
authors.initials: GA
authors.lastname: Maldarelli
authors.affiliation: Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Kurt H
authors.initials: KH
authors.lastname: Piepenbrink
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21201, USA.
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Scott
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21201, USA.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Freiberg
authors.affiliation: Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21201, USA.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Achermann
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21201, USA.
authors.forename: Robert K
authors.initials: RK
authors.lastname: Ernst
authors.affiliation: Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21201, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Shirtliff
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD 21201, USA Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Sundberg
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Donnenberg
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA Department of Veterans Affairs, VA Maryland Health Care System, Baltimore, MD 21201, USA evonrose@medicine.umaryland.edu.
authors.forename: Erik C
authors.initials: EC
authors.lastname: von Rosenvinge
mesh_terms: D018441:Biofilms / Q000254:growth & development*; D016360:Clostridioides difficile / Q000502:physiology*; D016000:Cluster Analysis; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology*; D036781:Fimbriae Proteins / Q000235:genetics; D010861:Fimbriae, Bacterial / Q000502:physiology*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009154:Mutation; D010641:Phenotype; D059467:Transcriptome
chemical_list: D036781:Fimbriae Proteins
keywords: Clostridium difficile; biofilm; type IV pili