Systematic analysis of the signal responsive gene regulatory network governing Myxococcus xanthus development

Studies of signal-induced gene expression in bacteria have contributed to understanding of how bacteria cope with environmental stress. As an extensively studied model, Myxococcus xanthus provides fascinating insights into how changes at the level of gene expression enable which bacteria to survive environmental insults such as nutrient limitation. Upon starvation M. xanthus cells glide into aggregates and form mounds that mature into fruiting bodies as some cells form spores. Previously, our group defined 24-30 h poststarvation as the critical period for commitment to spore formation, when cells commit to form spores despite perturbation of the starvation signal by nutrient addition. The process of multicellular development that culminates in sporulation is governed by a network of signal-responsive transcription factors that integrate signals for starvation and cellular alignment. In this dissertation I present the first systematic approach to elucidate the network dynamics during the commitment period.In the network, MrpC is a starvation-responsive transcription factor, whereas FruA is a transcription factor that responds to cellular alignment conveyed by C-signaling. Transcription of fruA is dependent on MrpC binding, and FruA activity is proposed to be posttranslationally regulated by C-signaling, although the mechanism is unknown. FruA and MrpC cooperatively regulate transcription of the dev operon. My systematic analysis of the network dynamics supported a model in which posttranslational activation of FruA by C-signaling is critical for dev transcription and for commitment to spore formation. Similar to dev, MrpC and C-signal-activated FruA combinatorially controlled transcription of the late-acting fadIJ operon involved in spore metabolism. Regulation of late-acting operons implicated in spore coat biogenesis (exoA-I, nfsA-H, MXAN_3259-MXAN_3263) was discovered to be under complex control by MrpC and FruA. My evidence suggests that transcription of these operons depends at least in part on a C-signal-dependent switch from negative regulation by unactivated FruA to positive regulation by activated FruA during the period leading up to and including commitment to sporulation. MrpC negatively regulated exo and MXAN_3259 during mound formation, but positively regulated nfs. During commitment to sporulation, MrpC continued to positively regulate nfs, switched to positive regulation of MXAN_3259, and continued to negatively regulate exo. A third transcription factor, Nla6, appeared to be a positive regulator of all the late genes. We propose that in combination with regulation by Nla6, differential regulation by FruA in response to C-signaling and by MrpC controls late gene expression to ensure that spore resistance and surface characteristics meet environmental demands.