Growth rate control of flagellar assembly in Escherichia coli strain RP437

Sim, Martin; Koirala, Santosh; Picton, David M; Strahl, Henrik; Hoskisson, Paul A.; Rao, Christopher V.; Gillespie, Colin S.; Aldridge, Phillip D.

doi:10.1038/srep41189

Cited by 34 publications

(37 citation statements)

References 39 publications

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“…This process was less efficient in cells grown on pyruvate and therefore could limit other physiological functions such as biofilm formation and motility. Indeed, the expression of the corresponding genes (e.g., dosC, ydaM and fliO, fliN and fliM) was reduced during E. coli growth on pyruvate and resembled their growth-ratedependent regulation observed in other studies (Sim et al 2017).…”

Section: Discussionsupporting

confidence: 78%

Reprogramming of gene expression in Escherichia coli cultured on pyruvate versus glucose

et al. 2019

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Previous studies revealed important roles of small RNAs (sRNAs) in regulation of bacterial metabolism, stress responses and virulence. However, only a minor fraction of sRNAs is well characterized with respect to the spectra of their targets, conditional expression profiles and actual mechanisms they use to regulate gene expression to control particular biological pathways. To learn more about the specific contribution of sRNAs to the global regulatory network controlling the Escherichia coli central carbon metabolism (CCM), we employed microarray analysis and compared transcriptome profiles of E. coli cells grown on two alternative minimal media supplemented with either pyruvate or glucose, respectively. Microarray analysis revealed that utilization of these alternative carbon sources led to profound differences in gene expression affecting all major gene clusters associated with CCM as well as expression of several known (CyaR, RyhB, GcvB and RyeA) and putative (C0652) sRNAs. To assess the impact of transcriptional reprogramming of gene expression on E. coli protein abundance, we also employed two-dimensional protein gel electrophoresis. Our experimental data made it possible to determine the major pathways for pyruvate assimilation when it is used as a sole carbon source and reveal the impact of other key processes (i.e., energy production, molecular transport and cell resistance to stress) associated with the CCM in E. coli. Moreover, some of these processes were apparently controlled by GcvB, RyhB and CyaR at the post-transcriptional level, thus indicating the complexity and interconnection of the regulatory networks that control CCM in bacteria.

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Section: Discussionsupporting

confidence: 78%

Reprogramming of gene expression in Escherichia coli cultured on pyruvate versus glucose

et al. 2019

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“…1C ). When this fluorescent protein fusion is expressed in cells, it forms spots associated with nascent C-rings that loosely correlate with the number of flagella 12 – 14 . By counting the number of spots per cell, we can determine the number of flagella made per cell.…”

Section: Resultsmentioning

confidence: 99%

“…FliM-GFP foci were quantified using Microbetracker on images captured using a Nikon Ti inverted microscope using filters and exposure times described previously 14 . Strains were grown to an OD600 of 0.5 to 0.6 and cells immobilised using a 1% agarose pad containing 10% LB 14 , 17 . For each strain a minimum of 5 fields of view were captured from 3 independent repeats.…”

Section: Methodsmentioning

confidence: 99%

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Driving the expression of the Salmonella enterica sv Typhimurium flagellum using flhDC from Escherichia coli results in key regulatory and cellular differences

Albanna

Sim

Hoskisson

et al. 2018

Sci Rep

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The flagellar systems of Escherichia coli and Salmonella enterica exhibit a significant level of genetic and functional synteny. Both systems are controlled by the flagellar specific master regulator FlhD4C2. Since the early days of genetic analyses of flagellar systems it has been known that E. coli flhDC can complement a ∆flhDC mutant in S. enterica. The genomic revolution has identified how genetic changes to transcription factors and/or DNA binding sites can impact the phenotypic outcome across related species. We were therefore interested in asking: using modern tools to interrogate flagellar gene expression and assembly, what would the impact be of replacing the flhDC coding sequences in S. enterica for the E. coli genes at the flhDC S. entercia chromosomal locus? We show that even though all strains created are motile, flagellar gene expression is measurably lower when flhDCEC are present. These changes can be attributed to the impact of FlhD4C2 DNA recognition and the protein-protein interactions required to generate a stable FlhD4C2 complex. Furthermore, our data suggests that in E. coli the internal flagellar FliT regulatory feedback loop has a marked difference with respect to output of the flagellar systems. We argue due diligence is required in making assumptions based on heterologous expression of regulators and that even systems showing significant synteny may not behave in exactly the same manner.

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“…We also note that the present analysis was limited to steady-state exponential growth. Others have previously explored the temporal dynamics of flagellar gene expression at single-cell resolution (5, 8, 34) and identified a number of transient phenomena that cannot be explained by the working model developed in the present study. Many factors are known to affect the dynamic response of flagellar gene expression.…”

Section: Discussionmentioning

confidence: 97%

Two tandem mechanisms control bimodal expression of the flagellar genes inSalmonella enterica

Wang

Koirala

Aldridge

et al. 2019

Preprint

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ABSTRACTFlagellar gene expression is bimodal in Salmonella enterica. Under certain growth conditions, some cells express the flagellar genes whereas others do not. This results in mixed populations of motile and non-motile cells. In the present study, we found that two independent mechanisms control bimodal expression of the flagellar genes. One was previously found to result from a double negative-feedback loop involving the flagellar regulators YdiV and FliZ. This feedback loop governs bimodal expression of class 2 genes. In this work, a second mechanism was found to govern bimodal expression of class 3 genes. In particular, class 3 gene expression is still bimodal even when class 2 gene expression is not. Using a combination of experimental and modeling approaches, we found that class 3 bimodalilty results from the σ28-FlgM developmental checkpoint.IMPORTANCEMany bacterial use flagella to swim in liquids and swarm over surface. In Salmonella enterica, over fifty genes are required to assemble flagella. The expression of these genes is tightly regulated. Previous studies have found that flagella gene expression is bimodal in S. enterica, which means that only a fraction of cells express flagellar genes and are motile. In the present study, we found that two separate mechanisms induce this bimodal response. One mechanism, which was previously identified, tunes the fraction of motile cells in response to nutrients. The other results from a developmental checkpoint that couples flagellar gene expression to flagellar assembly. Collectively, these results further our understanding of how flagellar gene expression is regulated in S. enterica.

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Growth rate control of flagellar assembly in Escherichia coli strain RP437

Cited by 34 publications

References 39 publications

Reprogramming of gene expression in Escherichia coli cultured on pyruvate versus glucose

Reprogramming of gene expression in Escherichia coli cultured on pyruvate versus glucose

Driving the expression of the Salmonella enterica sv Typhimurium flagellum using flhDC from Escherichia coli results in key regulatory and cellular differences

Two tandem mechanisms control bimodal expression of the flagellar genes inSalmonella enterica

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