An Electrostatic Interaction between BlpC and BlpH Dictates Pheromone Specificity in the Control of Bacteriocin Production and Immunity in Streptococcus pneumoniae

ABSTRACTTheblplocus ofStreptococcus pneumoniaesecretes and regulates bacteriocins, which mediate both intra- and interspecific competition in the human nasopharynx. There are four major alleles of the geneblpH, which encodes the receptor responsible for activating theblplocus when bound to one of four distinct peptide pheromones (BlpC). The allelic variation ofblpHis presumably explained by a need to restrict cross talk between competing strains. The BlpH protein sequences have polymorphisms distributed throughout the sequence, making identification of the peptide binding site difficult to predict. To identify the pheromone binding sites that dictate pheromone specificity, we have characterized the four major variants and two naturally occurring chimeric versions ofblpHin which recombination events appear to have joined two distinctblpHalleles together. Using these allelic variants, a series of laboratory-generated chimericblpHalleles, and site-directed mutants of both the receptor and peptide, we have demonstrated that BlpC binding to some BlpH types involves an electrostatic interaction between the oppositely charged residues of BlpC and the first transmembrane domain of BlpH. An additional recognition site was identified in the second extracellular loop. We identified naturally occurring BlpH types that have the capacity to respond to more than one BlpC type; however, this change in specificity results in a commensurate drop in overall sensitivity. These natural recombination events were presumably selected for to balance the need to sense bacteriocin-secreting neighbors with the need to turn on bacteriocin production at a low density.IMPORTANCEBacteria use quorum sensing to optimize gene expression to accommodate for local bacterial density and diffusion rates. To prevent interception of quorum-sensing signals by neighboring strains, the genomes of single species often encode strain-specific signal/receptor pairs. Theblplocus inStreptococcus pneumoniaethat drives bacteriocin secretion is controlled by quorum sensing that involves the interaction of the signal/receptor pair BlpC/BlpH. We show that the pneumococcal population can be divided into several distinct BlpC/BlpH pairs; however, there are examples of naturally occurring chimeric receptors that can bind to more than one BlpC type. The trade-off for this broadened specificity is a loss of overall receptor sensitivity. This suggests that under certain conditions, the advantage of signal interception can trump the requirements for self-induction.

Pheromonal Production of and Response to Optically Active Epoxydienes in Some Geometrid Moths (Lepidoptera: Geometridae)

In pheromone extracts of calling female Chiasma clathrata L. (Lepidoptera: Geometridae), a defoliator pest of alfalfa, (Z,Z,Z)-3,6,9-heptadecatriene and (Z,Z)-6,9-cis-3,4-epoxyheptadecadiene was identified. Chiral gas chromatography using a modified cyclodextrin and synthetic reference samples proved the natural epoxide to show (3R, 4S)-configuration. In field trapping tests, only the pure (3R, 4S)-enantiom er of the epoxide attracted males. The addition of the triene component was synergistic. Males of the sympatric species Tephrina arenacearia Hbn. (Lepidoptera: Geometridae) were caught only in traps with baits containing the (3S, 4R)-enantiomer [together with a previously described minor component, (Z,Z)-3,9-cis- 6,7-epoxyheptadecadiene]. In trapping tests conducted in a different biotope, Abraxas grossulariata L. (Lepidoptera: Geometridae) males were attracted by the (3S, 4R)-enantiomer, whereas the (3R, 4S)-enantiomer attracted a close relative, Abraxas sylvata Scop. (Lepidoptera: Geometridae). The present results suggest that one of the key mechanisms responsible for pheromone specificity among both the two alfalfa geometrids and the two A braxas species in their respective biotops, may be the use of different enantiomers of the same polyenederived epoxide as a sex pheromone component. It is probable that this discrimination mechanism is widespread among moth species utilizing epoxide pheromone components

Sexual Pheromones and Related Egg Secretions in Laminariales (Phaeophyta)

Volatile egg secretions of 15 species belonging to 5 families of the brown algal order Laminariales have been analyzed by high resolution glass capillary gas chromatography. 16 components have been identified and partially determined quantitatively. The composition of the pheromone bouquets and pheromone specificity for induction of spermatozoid-release and -chemotaxis are discussed in respect to ecophysiological functions and significance as chemo- taxonomic markers.