title: Differential expression of conserved protease genes in crucifer-attacking pathovars of Xanthomonas campestris.
abstract: Strains of Xanthomonas campestris pathovars armoraciae and raphani, which cause leaf spotting diseases in brassicas, produce a major extracellular protease in liquid culture which was partially purified. The protease (PRT 3) was a zinc-requiring metalloenzyme and was readily distinguishable from the two previously characterized proteases (PRT 1 and PRT 2) of X. campestris pv. campestris by the pattern of degradation of beta-casein and sensitivity to inhibitors. PRT 3 was produced at a low level in the vascular brassica pathogen X. campestris pv. campestris (five strains tested), in which PRT 1 and PRT 2 predominate. In contrast, expression of PRT 1, a serine protease, could not be detected in the six tested strains of the leaf spotting mesophyll pathogens. However, all these strains had DNA fragments which hybridized to a prtA probe and which probably carry a functional prtA (the structural gene for PRT 1). The structural gene for PRT 3 (prtC) was cloned by screening a genomic library of X. campestris pv. raphani in a protease-deficient X. campestris pv. campestris strain. Subcloning and Tn5 mutagenesis located the structural gene to 1.2 kb of DNA. DNA fragments which hybridized to the structural gene were found in all strains of the crucifer-attacking X. campestris pathovars tested as well as in a number of other pathovars. Experiments in which the pattern of protease production of the pathovars was manipulated by introduction of cloned genes into heterologous pathovars suggested that no determinative relationship exists between the pattern of protease gene expression and the (vascular or mesophyllic) mode of pathogenesis.
authors.affiliation: Sainsbury Laboratory, John Innes Centre for Plant Science Research, Norwich, United Kingdom.
authors.forename: J M
authors.initials: JM
authors.lastname: Dow
authors.forename: M J
authors.initials: MJ
authors.lastname: Fan
authors.forename: M A
authors.initials: MA
authors.lastname: Newman
authors.forename: M J
authors.initials: MJ
authors.lastname: Daniels
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000382:microbiology*; D003001:Cloning, Molecular / Q000379:methods; D010450:Endopeptidases / Q000032:analysis / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D013045:Species Specificity; D016959:Xanthomonas campestris / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity
chemical_list: D010450:Endopeptidases