title: The early stages of filamentous phage phiLf infection require the host transcription factor, Clp.
abstract: Xanthomonas campestris pv. campestris produces great amounts of an exopolysaccharide (EPS), xanthan gum. Eight eps loci involved in biosynthesis of the EPS were previously located in the chromosome map of strain Xc17. In this study, the eps8 region was cloned, sequenced and found to contain a crp homologue whose deduced amino acid sequence possesses similarity to that of the cyclic AMP receptor protein of bacteria, with the highest identity (97%) being shared with the X. campestris pv. campestris B-1459 clp gene previously shown to be involved in pathogenicity and regulation of the production of xanthan, extracellular enzymes, and pigment (de Crecy-Lagard V., Glaser P., Lejeune P., Sismeiro O., Barber C.E., Daniels M.J., and Danchin A., J. Bacteriol. 172:5877-5883, 1990). Based on sequence identity, pleiotropic effects of the mutation, the ability to complement an Escherichia coli cya crp mutant, and Southern hybridization detecting a single copy in the chromosome, we propose this eps8 gene to be the Xc17 clp. In addition to the previously reported properties, a clp mutant (AU56E) cannot be plaqued with filamentous phage phiLf, although it retains the capability to support phiLf DNA replication and release authentic phage particles upon electroporation of the RF DNA. Infective center assays demonstrated that the frequency of infection is 460- to 7,500-fold lower in AU56E compared to that in the wild-type Xc17. Electron microscopy, which showed no surface appendages other than the monotrichous flagellum, confirmed that AU56E drastically diminishes in the efficiency of phage adsorption. These results suggest Clp to be regulating the biosynthesis of the primary receptor, most likely a type IV pilus. Upstream to clp is a homologue of the E. coli speD gene required for spermidine synthesis. Mutation of the clp flanking regions and transcriptional analyses suggest clp to be monocistronic and the only gene contained at the eps8 locus.
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taiwan, Republic of China.
authors.forename: T C
authors.initials: TC
authors.lastname: Lee
authors.forename: S T
authors.initials: ST
authors.lastname: Chen
authors.forename: M C
authors.initials: MC
authors.lastname: Lee
authors.forename: C M
authors.initials: CM
authors.lastname: Chang
authors.forename: C H
authors.initials: CH
authors.lastname: Chen
authors.forename: S F
authors.initials: SF
authors.lastname: Weng
authors.forename: Y H
authors.initials: YH
authors.lastname: Tseng
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D017103:Bacillus Phages / Q000502:physiology*; D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016959:Xanthomonas campestris / Q000235:genetics / Q000502:physiology* / Q000821:virology*
chemical_list: D001426:Bacterial Proteins; C436751:Clp protein, Xanthomonas campestris; D011135:Polysaccharides, Bacterial; D014157:Transcription Factors; C002563:xanthan gum