Molecular characterization of the clumping factor (fibrinogen receptor) of Staphylococcus aureus

Four mutants of Staphylococcus aureus strain Newman that were defective in the fibrinogen receptor (clumping factor) were isolated by transposon Tn917 mutagenesis. Southern hybridization analysis of the mutants identified transposon-host DNA junction fragments, one of which was cloned and used to generate a probe to identify and clone the wild-type clumping factor locus (clfA). The mutants failed to form clumps in soluble fibrinogen and adhered poorly to polymethylmethacrylate (PMMA) coverslips coated with fibrinogen. A single copy of the clfA gene, when introduced into the chromosome of the mutant strains, fuily compiemented the ciumping deficiency of these strains and restored the ability of these mutants to adhere to fibrinogen-coated PMMA. in addition, the cloned clfA gene on a shuttle plasmid aiiowed the weakiy ciumping strain 8325-4 to form clumps with the same avidity as the wild-type strain Newman and also significantly enhanced the adherence of 8325-4 strains. Thus the formation of clumps in soluble fibrinogen correlated with adherence of bacteria to solid-phase fibrinogen. The clfA gene encodes a fibrinogen-binding protein with an apparent molecular mass of c. 130 kDa. The amino acid sequence of the protein was deduced from the DNA sequence; it was predicted that a 896 residue protein (molecular mass 92 kDa) would be expressed. The putative ClfA protein has features that suggest that it is associated with the ceil surface. Furthermore it contains a novel 308 residue region comprising dipeptide repeats predominantly of Asp and Ser ending 28 residues upstream from the LPXTG motif common to wall-associated proteins. Significant homology was found between the ClfA protein and the fibronectin-binding proteins of S. S. aureus, particularly in the N-and C-termini.     

Growth and reproduction of an estuarine population of the colonial hydroidCordylophora caspia (Pallas) in the northern Baltic Sea

Growth and reproduction of the colonial hydroidCordylophora caspia were monitored during the breeding season in natural conditions. In 1987, a life history study was carried out on the upright stems of the main stolon. Mean size of uprights varied cyclically. The first peak coincided with the peak number of sexual hydranths, after which the mean upright length decreased, possibly indicating somatic costs of sexual reproduction. Extrinsic factors like flooding may also have contributed to cyclical changes in upright size. In 1988 and 1989, colonies were reared on experimental plates in the estuary. In 1988, colonies grew until mid July, after which they regressed to a dormant condition and then started growing again in mid August. Predation and space competition are discussed as possible causes for this dormancy in the middle of the growing season. In 1989, colonies grew continually, with the exception of a decline in colony biomass and number of feeding hydranths at the end of July, just following the peak of sexual reproduction. Sexual reproduction started in the early stages of colonial development for all years. During early summer,C. caspia allocated resources simultaneously in colonial growth and sexual reproduction. However, sexual reproduction had a clear peak in mid summer, and thereafter sexual reproduction ceased while colonial growth continued.