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In this study we have demonstrated the activity of a choline phosphate cytidylyltransferase in cell free extracts of Streptococcus pneumoniae. Southern blot analysis of restricted S. pneumoniae genomic DNA probed with the gene coding for the choline phosphate cytidylyltransferase of Saccharomyces cerevisiae demonstrated that there is homology between the S. cerevisiae cct gene and genomic DNA of S. pneumoniae. We believe that this enzyme is involved in the biosynthesis of the choline containing cell wall antigens, teichoic acid and lipoteichoic acid, catalysing the activation of choline phosphate to CDP-choline which is then incorporated into the polysaccharide moiety.

The choline-containing teichoic and lipoteichoic acids play an important part in cell wall metabolism of Streptococcus pneumoniae. We propose that a choline kinase enzyme has a role in the synthesis of these antigens. The presence of this enzyme was demonstrated in cell free extracts of S. pneumoniae by measuring the fall in ATP concentration due to phosphorylation of choline. Genomic DNA of S. pneumoniae hybridised with a probe consisting of an internal fragment of the choline kinase gene of Saccharomyces cerevisiae and one consisting of the choline binding domain of lytA.

The effects of cold-shock on the resistance of plasmid-free and plasmid-carrying Escherichia coli to acrylate and copper ions have been tested. Such shock, produced by transfer from 37 to 5 degrees C, with 60 min incubation at the lower temperature, significantly enhanced the resistance of all the tested strains to both inhibitors. Such resistances may have arisen because the inhibitory agents are less able, due to porin changes, to penetrate into the organisms after cold-shock. It is more likely, however, that inhibitor penetration is unaffected but that cold-shocked organisms are better able to repair the damage caused (e.g. to membranes, DNA or cellular enzymes) by the inhibitors.