Identification of homologues to the pathogenicity factor Pat-1, a putative serine protease of Clavibacter michiganensis subsp. michiganensis

Hybridization of Clavibacter michiganensis subsp. michiganensis total DNA against the pathogenicity gene pat-1 indicated the presence of pat-1 homologous nucleotide sequences on the chromosome and on plasmid pCM2. Isolation of the corresponding DNA fragments and nucleotide sequence determination sho...

Full description

Saved in:

Bibliographic Details
Published in:	Microbiological research Vol. 160; no. 4; pp. 417 - 427
Main Authors:	Burger, Annette, Gräfen, Ines, Engemann, Jutta, Niermann, Erik, Pieper, Martina, Kirchner, Oliver, Gartemann, Karl-Heinz, Eichenlaub, Rudolf
Format:	Journal Article
Language:	English
Published:	Germany Elsevier GmbH 01-01-2005
Subjects:	Actinomycetales - enzymology Actinomycetales - genetics Actinomycetales - pathogenicity Amino Acid Motifs Amino Acid Sequence Bacterial Proteins - genetics Chromosomes, Bacterial - genetics Clavibacter Clavibacter michiganensis Clavibacter michiganensis subsp. michiganensis Conserved Sequence culture media DNA, Bacterial - chemistry DNA, Bacterial - genetics genes genotype hybridization Molecular Sequence Data Nucleic Acid Hybridization nucleotide sequences pathogenicity Phytopathogen plant pathogenic fungi plasmids Plasmids - genetics Protein Sorting Signals - genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Serine Endopeptidases - genetics Serine protease serine proteinases site-directed mutagenesis virulence Virulence - genetics Serine protease Clavibacter Phytopathogen
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Hybridization of Clavibacter michiganensis subsp. michiganensis total DNA against the pathogenicity gene pat-1 indicated the presence of pat-1 homologous nucleotide sequences on the chromosome and on plasmid pCM2. Isolation of the corresponding DNA fragments and nucleotide sequence determination showed that there are three pat-1 homologous genes: chpA (chromosome) and phpA and phpB (plasmid pCM2). The gene products share common characteristics, i.e. a signal sequence for Sec-dependent secretion, a serine protease motif, and six cysteine residues at conserved positions. Gene chpA located on the chromosome is a pseudogene since it contains a translational stop codon after 97 of 280 amino acids. In contrast to pat-1, cloning of the plasmid encoded homologs phpA and phpB into the avirulent plasmid free Cmm strain CMM100 did not result in a virulent phenotype. So far no proteolytic activity could be demonstrated for Pat-1, however, site specific mutagenesis of pat-1 showed that the serine residue in the motif GDSGG is required for the virulent phenotype of pat-1 and thus Pat-1 could be a functional protease.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0944-5013 1618-0623
DOI:	10.1016/j.micres.2005.03.006