title: A novel DNA-binding protein from Campylobacter jejuni bacteriophage NCTC12673.
abstract: We previously suggested that the double-stranded genomic DNA of Campylobacter jejuni bacteriophage NCTC12673 was complexed with proteins. Mass spectrometry of peptides obtained from tryptic digests of purified phage DNA indicated that phage protein Gp001 co-purified with the DNA. Gp001 is an acidic protein that lacks any obvious homology or conserved domains found in known DNA-binding proteins. The DNA-binding ability of recombinant Gp001 was examined using an electrophoretic mobility shift assay. Slow DNA-Gp001 complex formation was observed at pH 5.5, but not at neutral or basic pH. This nucleoprotein complex had difficulty entering agarose gels used in the assay while proteinase K pretreatment released the DNA from the complex. No mobility shift was observed when the DNA was immediately subjected to electrophoresis after mixing with Gp001, even if both components were separately pre-incubated at pH 5.5. The complexed DNA was unable to transform chemically competent Escherichia coli cells and was less susceptible to degradation by nucleases. The formation of Gp001-DNA complexes at low pH may provide a mechanism for maintaining DNA integrity while the phage pursues its host through the gastrointestinal tract. Also, this feature can potentially be used to improve DNA delivery protocols applied in gene therapy.
authors.affiliation: Department of Biological Sciences and Alberta Glycomics Centre, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Denis
authors.initials: D
authors.lastname: Arutyunov
authors.affiliation: Department of Biological Sciences and Alberta Glycomics Centre, University of Alberta, Edmonton, Alberta, Canada cszymans@ualberta.ca.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Szymanski
mesh_terms: D001435:Bacteriophages / Q000737:chemistry* / Q000235:genetics*; D016123:Campylobacter jejuni / Q000821:virology*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins
keywords: Campylobacter jejuni; DNA-binding protein; bacteriophage