Biology subjects

Higgs, P. I.

Publications and source records attributed to Higgs, P. I..

4 recordsLinked to original sources

bioRxiv2026

The orphan histidine kinase TodK controls Myxococcus xanthus biofilm development by inactivating the CRP/Fnr homolog, MrpC

Environmental bacteria have abundant signaling systems wired into complex gene regulatory networks to adapt to fluctuating conditions. In Myxococcus xanthus, starvation triggers a developmental program (specialized biofilm) that produces spore-filled multicellular fruiting bodies surrounded by a distinct quiescent state termed peripheral rods. Fruiting body structure as well as the proportion of cells following each fate can be tuned by a large repertoire of signaling proteins, including numerous orphan histidine kinases. Here, we focus on the histidine kinase TodK which was previously demonstrated to influence developmental progression. We find that loss of TodK produces distinct developmental phenotypes that vary with environmental conditions. To quantify these effects, we developed an image-analysis pipeline that measures aggregation and fruiting body patterning during development on nutrient-limited agar. These analyses revealed the todK mutant precociously aggregates particularly at the peripheries of the colony. Under submerged-culture conditions, initial production of aggregates was not accelerated but aggregates exhibited accelerated progression to mature fruiting bodies. Overexpression of active TodK completely blocked fruiting body formation. Molecular analyses demonstrated that TodK overproduction suppressed expression of core developmental regulators including FruA and CsgA (C-signal). Interestingly, protein accumulation of MrpC, necessary for expression of both FruA and the C-signal was not significantly perturbed suggesting TodK silences MrpC transcriptional activity. Together, these findings establish TodK as a modulator of developmental progression and demonstrate how quantitative phenotyping approaches can reveal biologically meaningful functions for orphan histidine kinases whose mutant phenotypes might otherwise appear subtle. Summary StatementQuantitative analysis of multicellular development reveals previously hidden functions of an orphan histidine kinase, highlighting the importance of robust phenotyping approaches for understanding bacterial signaling networks.

microbiology↗

bioRxiv2019

An amino-terminal threonine/serine motif is necessary for activity of the Crp/Fnr homolog, MrpC, and for Myxococcus xanthus developmental robustness

The Crp/Fnr family of transcriptional regulators play central roles in transcriptional control of diverse physiological responses. Activation of individual family members is controlled by a surprising diversity of mechanisms tuned to the particular physiological responses or lifestyles that they regulate. MrpC is a Crp/Fnr homolog that plays an essential role in controlling the Myxococcus xanthus developmental program. A long-standing model proposed that MrpC activity is controlled by the Pkn8/Pkn14 serine/threonine kinase cascade which phosphorylates MrpC on threonine residue(s) located in its extreme amino terminus. In this study, we demonstrate that a stretch of consecutive threonine and serine residues, T21 T22 S23 S24, is necessary for MrpC activity by promoting efficient DNA binding. Mass spectrometry analysis indicated the TTSS motif is not directly phosphorylated by Pkn14 in vitro but is necessary for efficient Pkn14-dependent phosphorylation on several residues in the remainder of the protein. Pkn8 and Pkn14 kinase activities do not play obvious roles in controlling MrpC activity in wild type M. xanthus under laboratory conditions, but likely modulate MrpC DNA binding in response to unknown environmental conditions. Interestingly, mutational analysis of the TTSS motif caused non-robust developmental phenotypes, revealing that MrpC plays a role in developmental buffering.

microbiology↗

bioRxiv2019

A negative autoregulation network motif is required for synchronized Myxococcus xanthus development

Transcription factor autoregulation is a simple network motif (recurring circuit) built into genetic regulatory networks that direct cell behavior. Negative autoregulation (NAR) network motifs are particularly abundant in bacteria and provide specific functions, such as buffering against transcriptional noise. Here, we investigate the phenotypic consequence of perturbing NAR of a major transcription factor, MrpC, that controls the multicellular development program of the bacterium Myxococcus xanthus. Launch of the developmental program directs certain cells in the population to first aggregate into haystack-shaped mounds, and then to differentiate into environmentally resistant spores to form mature fruiting bodies. Perturbation of MrpC NAR causes a striking phenotype in which cells lose synchronized transition from aggregation to sporulation. Instead, some cells abruptly exit aggregation centers and remain locked in a cohesive swarming state, while the remaining cells transition to spores inside residual fruiting bodies. As predicted, disruption of MrpC NAR led to an increased and broadened population distribution of mrpC expression. Examination of MrpC levels in developmental subpopulations during in situ development demonstrated cells locked in the swarms contained intermediate MrpC levels insufficient to promote sporulation. These results suggest an inherent property of NAR motifs that function in multicellular developmental programs is to facilitate synchronized responses. Significance StatementAll organisms use regulatory networks for cellular homeostasis, mediating appropriate responses to environmental changes, or to direct animal development. Understanding how the basic building blocks (motifs) of regulatory networks contribute to these processes is essential to mitigate the effects of mutations in regulatory networks (i.e. cancers) or to synthesize beneficial organisms. In this study, we demonstrate that a common regulatory motif, a transcription factor that represses its own expression, helps synchronize cells that engage in collective behaviors.

developmental biology↗

bioRxiv2019

Global transcriptome analysis of the Myxococcus xanthus multicellular developmental program

The bacteria Myxococcus xanthus exhibit a complex multicellular life cycle. In the presence of nutrients, cells prey cooperatively. Upon starvation, they enter a developmental cycle wherein cells aggregate to produce macroscopic fruiting bodies filled with resistant myxospores. We used RNA-Seq technology to examine the global transcriptome of the 96 h developmental program. This data revealed that many genes were sequentially expressed in discrete modules, with expression peaking during aggregation, in the transition from aggregation to sporulation, or during sporulation. Analysis of genes expressed at each specific time point provided a global framework integrating regulatory factors coordinating motility and differentiation in the developmental program. These data provided insights as to how starving cells obtain energy and precursors necessary for assembly of fruiting bodies and into developmental production of secondary metabolites. This study offers the first global view of developmental transcriptional profiles and provides an important scaffold for future studies. IMPACT STATEMENTInvestigation of global gene expression profiles during formation of the Myxococcus xanthus specialized biofilm reveals a genetic regulatory network that coordinates cell motility, differentiation, and secondary metabolite production.

microbiology↗