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• W2918102758 abstract "Brachyspira (B.) hyodysenteriae is the etiological agent of swine dysentery, a mucohaemorrhagic diarrhea causing significant economic losses. The pathogenesis of SD is still not fully understood. The hypothesis initiating this work was that the B. hyodysenteriae genome, in comparison to the other four Brachyspira species occuring in swine, possesses species-specific genes which might be responsible for its unique pathogenic properties. Based on this hypothesis a Representational Difference Analysis (RDA) was performed using B. hyodysenteriae B204 as 'tester' and the combined DNA from the reference strains of the other four Brachyspira species as 'driver'. Two B. hyodysenteriae-specific DNA fragments designated as BH100 and BH400 were identified; they hybridized to different locations on the genome of type strain B. hyodysenteriae B78T and are present in one copy only. Their species-specificity was confirmed by Southern blot analyses with Brachyspira ssp. reference strains and field isolates. One RDA fragment is part of a 2130 bp ORF designated as bhoA (B. hyodysenteriae orphan protein A). Database search revealed limited homology to sensor transduction kinases of Dictyostelium discoideum. Based on the second RDA fragment a putative fructose-specific phosphotransferase system (PTS) of B. hyodysenteriae was identified consisting of genes encoding a putative transcriptional repressor (fruR), PTS enzymes IIA (fruA) and IIBC (fruBC), and fructose-bisphosphate aldolase (fbaA). Nucleotide sequence analysis reavealed direct repeats between the fruR and fruA genes which might serve as operator, and inverted repeats downstream of the fbaA gene possibly representing intrinsic transcriptional terminators. Western blot and Northern blot analyses revealed a fructose-induced upregulation of transcription and translation of the PTS genes fruA and fruBC, respectively, and a constitutive transcription and translation of the fruR gene. In addition, Northern blot analysis revealed several transcripts reacting with a fruBC-derived probe indicating a posttranscriptional processing of the polycistronic mRNA. In combination with the result of a primer extension analysis a negative regulation of the B. hyodysenteriae-specific PTS with posttranscriptional cleavage of the mRNA is proposed. A chemotaxis assay revealed chemotaxis of B. hyodysenteriae towards fructose. As PTSs are involved in positive chemotaxis towards carbohydrates, the fructose-specific PTS identified in B. hyodysenteriae might play a role in chemotaxis towards fructose; a potential role in pathogenesis of SD remains to be investigated." @default.
• W2918102758 created "2019-03-02" @default.
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• W2918102758 date "2003-01-01" @default.
• W2918102758 modified "2023-09-27" @default.
• W2918102758 title "A representational difference analysis and the identification of a fructose-specific phosphotransferase system of Brachyspira hyodysenteriae" @default.
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