It has long been hypothesized that subcellular positioning of chromosomal loci in bacteria may be influenced by gene function and expression state. Here we provide direct evidence membrane protein affects the position Escherichia coli. For two different proteins, observed a dramatic shift their genetic toward upon induction. In related systems which cytoplasmic was produced, or translation eliminated mutating start codon, not observed. Antibiotics block transcription similarly prevented...

Most bacteria contain both eukaryotic-like Ser/Thr kinases (eSTKs) and phosphatases (eSTPs). Their role in bacterial physiology is not currently well understood large part because the conditions where eSTKs are active generally known. However, all sequenced Gram-positive have a highly conserved eSTK with extracellular PASTA repeats that bind cell wall derived muropeptides. Here, we report bacterium Bacillus subtilis, PASTA-containing PrkC its cognate eSTP PrpC converge essential WalRK...

Abstract β‐Lactams represent one of the most important classes antibiotics discovered to date. These agents block Lipid II processing and cell wall biosynthesis through inactivation penicillin‐binding proteins (PBPs). PBPs enzymatically load building blocks from carrier molecules onto growing scaffold during growth division. II, a bottleneck in biosynthesis, is target some potent clinical use. Despite immense therapeutic value this biosynthetic pathway, PBP–Lipid association has not been...

Abstract Isogenic populations of cells exhibit phenotypic variability that has specific physiological consequences. Individual bacteria within a population can differ in antibiotic tolerance, but whether this be regulated or is generally an unavoidable consequence stochastic fluctuations unclear. Here we report gene encoding bacterial (p)ppGpp synthetase Bacillus subtilis , sasA exhibits high levels extrinsic noise expression. We find by multisite phosphorylation the transcription factor...

Abstract β‐Lactams represent one of the most important classes antibiotics discovered to date. These agents block Lipid II processing and cell wall biosynthesis through inactivation penicillin‐binding proteins (PBPs). PBPs enzymatically load building blocks from carrier molecules onto growing scaffold during growth division. II, a bottleneck in biosynthesis, is target some potent clinical use. Despite immense therapeutic value this biosynthetic pathway, PBP–Lipid association has not been...

Previously, an unexplained subcellular localization was reported for a functional fluorescent protein fusion to the response regulator OmpR in Escherichia coli. The pronounced regions of increased fluorescence, or foci, are dependent on phosphorylation and do not occupy fixed, easily identifiable positions, such as poles mid-cell. Here we show that foci due OmpR-YFP (yellow protein) binding specific sites chromosome. To identify positions quantify their fluorescence intensity, used simple...

At the single-cell level, protein kinase activity is typically inferred from downstream transcriptional reporters. However, promoters are often coregulated by several pathways, making of a specific difficult to deconvolve. Here, we present modular, direct, and sensors bacterial activity, including FRET-based sensors, as well synthetic transcription factor based on lactose repressor (LacI) that has been engineered respond phosphorylation. We demonstrate utility these in measuring PrkC,...

Abstract Isogenic populations of cells exhibit phenotypic variability that has specific physiological consequences. For example, individual bacteria within a population can differ in their sensitivity to an antibiotic, but whether this be regulated or is generally unavoidable consequence stochastic fluctuations unclear. We observed bacterial stress response gene, the (p)ppGpp synthetase sasA , exhibits high levels extrinsic noise expression, suggestive regulatory process. traced convergence...

Abstract A lack of direct single-cell readouts for bacterial kinase activity remains a major barrier to our understanding most signaling systems. At the single-cell-level, protein is typically inferred by downstream transcriptional reporters. Complicating this approach in vivo , promoters are often co-regulated several pathways, making specific difficult deconvolve. Here, we have designed and constructed new, sensors activity, including FRET-based sensors, as well synthetic transcription...