Identification of a Differentially Expressed Oligopeptide Binding Protein (OppA2) in Streptococcus Uberis by Representational Difference Analysis of CDNA

Streptococcus uberis is an increasingly significant cause of intramammary infection in the dairy cow, presently responsible for approximately 33% of all cases of bovine mastitis in the United Kingdom. Following experimentally induced infection of the lactating mammary gland, S. uberis is found predominantly in the luminal areas of secretory alveoli and ductular tissue, indicating that much of the bacterial growth occurs in residual and newly synthesized milk. With the objective of identifying potential virulence determinants in a clinical isolate of S. uberis, we have used representational difference analysis of cDNA to identify genes that show modified expression in milk. We have identified a number of differentially expressed genes that may contribute to the overall pathogenicity of the organism. Of these, a transcript encoding a putative oligopeptide binding protein (OppA) was further characterized. We have found that S. uberis possesses two oppA-like open reading frames, oppA1 and oppA2, which are up-regulated to different degrees following growth in milk. Mutants lacking either oppA1 or oppA2 are viable and have an increased resistance to the toxic peptide derivative aminopterin; however, only mutants lacking oppA1 display a lower rate of growth in milk. In addition, expression of the oppA genes appears to be coordinated by different mechanisms. We conclude that the oppA genes encode oligopeptide binding proteins, possibly displaying different specificities, required for the efficient growth of S. uberis in milk.