Project description:Transcriptional profiling of Myxococcus xanthus comparing wild-type and mutant gene expression at different developmental time points The mutants in our study, nla4, nla18, nla6 and nla28 are strains that carry a mutation in the corresponding genes nla4 (MXAN2516), nla18 (MXAN3692), nla6(MXAN4042), and nla28(MXAN1167). (MXAN numbers are the conventional gene names; nlas are common names). All four of these genes are response regulators of the ntrC family of transcriptional activators. The mutations were made by insertion of a plasmid (pCR2.1TOPO, invitrogen)into the particular activator gene. Plasmids used carry a region of homology (usually 350-500 bp in length) to the sequence of the activator gene coresponding to the highly conserved central domain that is characteristic of these ntrC family of activators. Plasmids were inserted into the chromosome via homologous recombination at the site of homology to the activator gene. These for genes were originally identified because their mutants have phenotypes with strong defects during development. In all four cases, a mutation in the particular gene gives a phenotype with incomplete or abolished development. In this study, the effects of these mutants were examined on developmental gene expression. Two condition experiment, normal wild-type vs mutant
Project description:In response to starvation Myxococcus xanthus initiates a developmental program that culminates in the formation of fruiting bodies inside which the rod-shaped cells differentiate to spores. Fruiting body formation depends on intercellular communication and two intercellular signals are known, the A-signal and the C-signal. Five genes have been identified which are required for A-signal synthesis. To begin to understand the function of the genes required for A-signal synthesis, we have analysed gene expression in the asgA and the asgB mutant. Keywords: Vegetative cells of WT (DK1622) and AsgA mutant (DK5057) and AsgB mutant (DK4398)
Project description:In Myxococcus xanthus 55% of the more than 250 two-component signal transduction systems (TCS) genes are orphan. We hypothesized that the histidine kinase SgmT and the response regulator DigR, which comprises a DNA binding domain of the HTH_Xer type, function together to regulate gene expression. We performed genome-wide expression profiling experiments to determine wether the same set of genes are differentially expressed in the ΔdigR and ΔsgmT mutants.
Project description:Myxococcus xanthus is a model organism for studying social behaviors and cell differentiation in bacteria. Upon nutrient depletion, M. xanthus cells initiate a developmental program that culminates in formation of spore-filled fruiting bodies and peripheral rods outside of fruiting bodies. Completion of this developmental program depends on fine-tuned spatial and temporal regulation of gene expression, intercellular communication, signaling by nucleotide-based second messengers, and motility. In order to understand stage-specific gene expression during growth and development, we extracted total RNA from vegetative cells (referred as 0 h of development) and from cells developed for 6, 12, 18 and 24 h under submerged conditions in two replicates.
Project description:Our ChipSeq analysis show that while FrzCD does not bind DNA specific regions, ParB binds parS consistent with what as been previously shown. The FrzCD chemoreceptor from the gliding bacterium Myxococcus xanthus forms cytoplasmic clusters that occupy a large central region of the cell body also occupied by the nucleoid. FrzCD directly binds to the nucleoid and the FrzCD binding to the DNA leads to the formation of chemosensory complexes. This supra-molecular organization is required for cooperative interactions between clustered receptors, in turn important for the modulation of bacterial social behaviors.