Characterization of the fimA gene encoding bundle-forming fimbriae of the plant pathogen Xanthomonas campestris pv. vesicatoria.
The fimA gene of Xanthomonas campestris pv. vesicatoria was identified and characterized. A 20-mer degenerate oligonucleotide complementary to the N-terminal amino acid sequence of the purified 15.5-kDa fimbrillin was used to locate fimA on a 2.6-kb SalI fragment of the X. campestris pv. vesicatoria 3240 genome. The nucleotide sequence of a 1.4-kb fragment containing the fimA region revealed two open reading frames predicting highly homologous proteins FimA and FimB. FimA, which was composed of 136 amino acids and had a calculated molecular weight of 14,302, showed high sequence identity to the type IV fimbrillin precursors. fimB predicted a protein product of 135 amino acids and a molecular weight of 13,854. The open reading frame for fimB contained near the 5' end a palindromic sequence with a terminator loop potential, and the expression level of fimB in vitro and in Xanthomonas was considerably lower than that of fimA. We detected an efficiently transcribed fimA-specific mRNA of 600 bases as well as two weakly expressed, longer mRNA species that reacted with both fimA and fimB. A homolog of fimA but not of fimB was detected by Southern hybridization in strains of X. campestris pv. vesicatoria, campestris, begoniae, translucens, and graminis. A fimA::omega mutant of strain 3240 was not significantly reduced in virulence or adhesiveness to tomato leaves. However, the fimA mutant was dramatically reduced in cell aggregation in laboratory cultures and on infected tomato leaves. The fimA mutant strain also exhibited decreased tolerance to UV light.
Ojanen-Reuhs
T
T
Department of Biosciences, University of Helsinki, Finland.
Kalkkinen
N
N
Westerlund-Wikström
B
B
van Doorn
J
J
Haahtela
K
K
Nurmiaho-Lassila
E L
EL
Wengelnik
K
K
Bonas
U
U
Korhonen
T K
TK
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D019426:Integrases; D018551:Solanum lycopersicum / Q000382:microbiology; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D010641:Phenotype; D018515:Plant Leaves / Q000382:microbiology; D016415:Sequence Alignment; D014466:Ultraviolet Rays; D016959:Xanthomonas campestris / Q000235:genetics* / Q000502:physiology / Q000528:radiation effects / Q000648:ultrastructure
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C081956:fimB protein, E coli; C063055:fimbrillin; D036781:Fimbriae Proteins; D019426:Integrases