Selection for ancient periodic motifs that do not impart DNA bending.

A ~10-11 bp periodicity in dinucleotides imparting DNA bending, with shorter periods found in organisms with positively-supercoiled DNA and longer periods found in organisms with negatively-supercoiled DNA, was previously suggested to assist in DNA compaction. However, when measured with more robust methods, variation in the observed periods between organisms with different growth temperatures is not consistent with that hypothesis. We demonstrate that dinucleotide periodicity does not arise solely by mutational biases but is under selection. We found variation between genomes in both the period and the suite of dinucleotides that are periodic. Whereas organisms with similar growth temperatures have highly variable periods, differences in periods increase with phylogenetic distance between organisms. In addition, while the suites of dinucleotides under selection for periodicity become more dissimilar among more distantly-related organisms, there is a core set of dinucleotides that are strongly periodic among genomes in all domains of life. Notably, this core set of periodic motifs are not involved in DNA bending. These data indicate that dinucleotide periodicity is an ancient genomic architecture which may play a role in shaping the evolution of genes and genomes.

The O-antigen mediates differential survival of Salmonella against communities of natural predators.

Antigenically distinct members of bacterial species can be differentially distributed in the environment. Predators known to consume antigenically distinct prey with different efficiencies are also differentially distributed. Here we show that antigenically distinct, but otherwise isogenic and physiologically indistinct, strains of Salmonella enterica show differential survival in natural soil, sediment and intestinal environments, where they would face a community of predators. Decline in overall cell numbers is attenuated by factors that inhibit the action of predators, including heat and antiprotozoal and antihelminthic drugs. Moreover, the fitness of strains facing these predators - calculated by comparing survival with and without treatments attenuating predator activity - varies between environments. These results suggest that relative survival in natural environments is arbitrated by communities of natural predators whose feeding preferences, if not species composition, vary between environments. These data support the hypothesis that survival against natural predators may drive the differential distribution of bacteria among microenvironments.

A likelihood approach to classifying fluorescent events collected by multicolor flow cytometry.

Flow cytometry is an effective tool for enumerating fluorescently-labeled microbes recovered from natural environments. However, low signal strength and the presence of fluorescent, non-cellular particles complicate the separation of cellular events from noise. Existing classification methods rely on the arbitrary placement of noise thresholds, resulting in potentially high rates of misclassification of fluorescent cells, thus precluding the robust estimation of the proportions of classes of fluorescent cells. Here we present a method for objectively separating signal from noise. Rather than setting an arbitrary noise threshold, the Z-scoring approach uses the Gaussian distribution of signal strength (a) to locate noise threshold for individual fluorophores, (b) to predict the likelihood of different fluorescent genotypes in producing the signal observed, and (c) to normalize the fraction of cellular events count for each fluorescent cell class. The likelihood framework allows rejection of alternative genotypes, leading to robust and reliable classification of fluorescent cells. Use of Z-scoring in classification of cells expressing multiple fluorophores, use of spillover in actively scoring events, and the successful classification of multiple fluorophores using a single detector within a flow cytometer are discussed. A software package that performs Z-scoring for cells labeled with one or more fluorophores is described.