title: Identification and initial characterisation of a protein involved in Campylobacter jejuni cell shape.
abstract: Campylobacter jejuni is the leading cause of bacterial food borne illness. While helical cell shape is considered important for C. jejuni pathogenesis, this bacterium is capable of adopting other morphologies. To better understand how helical-shaped C. jejuni maintain their shape and thus any associated colonisation, pathogenicity or other advantage, it is first important to identify the genes and proteins involved. So far, two peptidoglycan modifying enzymes Pgp1 and Pgp2 have been shown to be required for C. jejuni helical cell shape. We performed a visual screen of ∼2000 transposon mutants of C. jejuni for cell shape mutants. Whole genome sequence data of the mutants with altered cell shape, directed mutants, wild type stocks and isolated helical and rod-shaped 'wild type' C. jejuni, identified a number of different mutations in pgp1 and pgp2, which result in a change in helical to rod bacterial cell shape. We also identified an isolate with a loss of curvature. In this study, we have identified the genomic change in this isolate, and found that targeted deletion of the gene with the change resulted in bacteria with loss of curvature. Helical cell shape was restored by supplying the gene in trans. We examined the effect of loss of the gene on bacterial motility, adhesion and invasion of tissue culture cells and chicken colonisation, as well as the effect on the muropeptide profile of the peptidoglycan sacculus. Our work identifies another factor involved in helical cell shape.
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, UK.
authors.forename: Diane
authors.initials: D
authors.lastname: Esson
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, UK.
authors.forename: Srishti
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, New Museums Site, Pembroke Street, Cambridge, UK.
authors.forename: David
authors.initials: D
authors.lastname: Bailey
authors.affiliation: Department of Infection Biology, Institute for Infection and Global Health and School of Veterinary Science, University of Liverpool, Leahurst Campus, Neston, Cheshire, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Wigley
authors.affiliation: Department of Infection Biology, Institute for Infection and Global Health and School of Veterinary Science, University of Liverpool, Leahurst Campus, Neston, Cheshire, UK.
authors.forename: Amy
authors.initials: A
authors.lastname: Wedley
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors.forename: Alison E
authors.initials: AE
authors.lastname: Mather
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Méric
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, UK.
authors.forename: Pietro
authors.initials: P
authors.lastname: Mastroeni
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, UK.
authors.forename: Samuel K
authors.initials: SK
authors.lastname: Sheppard
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK; The London School of Hygiene and Tropical Medicine, London, UK.
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Thomson
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors.forename: Julian
authors.initials: J
authors.lastname: Parkhill
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, UK.
authors.forename: Duncan J
authors.initials: DJ
authors.lastname: Maskell
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, New Museums Site, Pembroke Street, Cambridge, UK.
authors.forename: Graham
authors.initials: G
authors.lastname: Christie
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge, UK. Electronic address: ajg60@cam.ac.uk.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Grant
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D018938:Caco-2 Cells; D016123:Campylobacter jejuni / Q000166:cytology* / Q000235:genetics* / Q000502:physiology; D002473:Cell Wall / Q000378:metabolism; D004251:DNA Transposable Elements; D004705:Endocytosis; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D008124:Locomotion; D016254:Mutagenesis, Insertional; D010457:Peptidoglycan / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D010457:Peptidoglycan
keywords: Campylobacter jejuni; Cell shape