A diverse set of prophage-mediated mechanisms protecting bacterial hosts from infection has been recently uncovered within cluster N mycobacteriophages isolated on the host, mc155. In that context, we unveil a novel defense mechanism in cluster N prophage Butters. By using bioinformatics analyses, phage plating efficiency experiments, microscopy, and immunoprecipitation assays, we show that Butters genes located in the central region of the genome play a key role in the defense against heterotypic viral attack.
View Article and Find Full Text PDFThe ability to gain quantifiable, single-cell data from time-lapse microscopy images is dependent upon cell segmentation and tracking. Here, we present a detailed protocol for obtaining quality time-lapse movies and introduce a method to identify (segment) and track cells based on machine learning techniques (Fiji's Trainable Weka Segmentation) and custom, open-source Python scripts. To provide a hands-on experience, we provide datasets obtained using the aforementioned protocol.
View Article and Find Full Text PDFIn cells, specific regulators often compete for limited amounts of a core enzymatic resource. It is typically assumed that competition leads to partitioning of core enzyme molecules among regulators at constant levels. Alternatively, however, different regulatory species could time share, or take turns utilizing, the core resource.
View Article and Find Full Text PDFThe correct functioning of cells requires the orchestration of multiple cellular processes, many of which are inherently dynamical. The conditions under which these dynamical processes entrain each other remain unclear. Here we use synthetic biology to address this question in the case of concurrent cellular oscillations.
View Article and Find Full Text PDFGene regulatory circuits can receive multiple simultaneous inputs, which can enter the system through different locations. It is thus necessary to establish how these genetic circuits integrate multiple inputs as a function of their relative entry points. Here, we use the dynamic circuit regulating competence for DNA uptake in Bacillus subtilis as a model system to investigate this issue.
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