A mathematical model of flagellar gene regulation and construction in Salmonella enterica

AbstractMillions of people worldwide develop foodborne illnesses caused by Salmonella enterica (S. enterica) every year. The pathogenesis of S. enterica depends on flagella, which are appendages that the bacteria use to move through the environment. Interestingly, populations of genetically identical bacteria exhibit heterogeneity in the number of flagella. To understand this heterogeneity and the regulation of flagella quantity, we propose a mathematical model that connects the flagellar gene regulatory network to flagellar construction. A regulatory network involving more than 60 genes controls flagellar assembly. The most important member of the network is the master operon, flhDC, which encodes the FlhD4C2 protein. FlhD4C2 controls the construction of flagella by initiating the production of hook basal bodies (HBBs), protein structures that anchor the flagella to the bacterium. By connecting a model of FlhD4C2 regulation to a model of HBB construction, we investigate the roles of various feedback mechanisms. Analysis of our model suggests that a combination of regulatory mechanisms at the protein and transcriptional levels induce bistable FlhD4C2 levels and heterogeneous numbers of flagella. Also, the balance of regulatory mechanisms that become active following HBB construction is sufficient to provide a counting mechanism for controlling the total number of flagella produced.Author summarySalmonella causes foodborne illnesses in millions of people worldwide each year. Flagella, which are appendages that the bacteria use to move through the environment, are a key factor in the infection process. Populations of genetically identical bacteria have been observed to contain both motile cells, generally with 6–10 flagella, and nonmotile cells, with no flagella. In this paper, we use mathematical models of the gene network that regulates flagellar construction to explore how the bacteria controls the number of flagella produced. We suggest that a bacterium must accumulate a threshold amount of a master regulator protein to initiate flagella production and failure to reach the threshold results in no flagella. Downstream mechanisms that impact the amount of master regulator protein are sufficient to determine how many flagella are produced.

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FlhF and Its GTPase Activity Are Required for Distinct Processes in Flagellar Gene Regulation and Biosynthesis in Campylobacter jejuni

Journal of Bacteriology ◽

10.1128/jb.00884-09 ◽

2009 ◽

Vol 191 (21) ◽

pp. 6602-6611 ◽

Cited By ~ 39

Author(s):

Murat Balaban ◽

Stephanie N. Joslin ◽

David R. Hendrixson

Keyword(s):

Gene Expression ◽

Gene Regulation ◽

Campylobacter Jejuni ◽

Regulatory System ◽

Flagellar Gene ◽

Gtpase Activity ◽

Gtp Hydrolysis ◽

Genetic Analyses ◽

Flagellar Biosynthesis ◽

Two Component

ABSTRACT FlhF proteins are putative GTPases that are often necessary for one or more steps in flagellar organelle development in polarly flagellated bacteria. In Campylobacter jejuni, FlhF is required for σ54-dependent flagellar gene expression and flagellar biosynthesis, but how FlhF influences these processes is unknown. Furthermore, the GTPase activity of any FlhF protein and the requirement of this speculated activity for steps in flagellar biosynthesis remain uncharacterized. We show here that C. jejuni FlhF hydrolyzes GTP, indicating that these proteins are GTPases. C. jejuni mutants producing FlhF proteins with reduced GTPase activity were not severely defective for σ54-dependent flagellar gene expression, unlike a mutant lacking FlhF. Instead, these mutants had a propensity to lack flagella or produce flagella in improper numbers or at nonpolar locations, indicating that GTP hydrolysis by FlhF is required for proper flagellar biosynthesis. Additional studies focused on elucidating a possible role for FlhF in σ54-dependent flagellar gene expression were conducted. These studies revealed that FlhF does not influence production of or signaling between the flagellar export apparatus and the FlgSR two-component regulatory system to activate σ54. Instead, our data suggest that FlhF functions in an independent pathway that converges with or works downstream of the flagellar export apparatus-FlgSR pathway to influence σ54-dependent gene expression. This study provides corroborative biochemical and genetic analyses suggesting that different activities of the C. jejuni FlhF GTPase are required for distinct steps in flagellar gene expression and biosynthesis. Our findings are likely applicable to many polarly flagellated bacteria that utilize FlhF in flagellar biosynthesis processes.

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Mathematical Modeling of the λ Switch

Handbook of Research on Computational Methodologies in Gene Regulatory Networks ◽

10.4018/978-1-60566-685-3.ch024 ◽

2010 ◽

pp. 573-603

Author(s):

Dmitriy Laschov ◽

Michael Margaliot

Keyword(s):

Mathematical Modeling ◽

Differential Equation ◽

Mathematical Model ◽

Gene Regulation ◽

Equilibrium Points ◽

Domain Of Attraction ◽

Verbal Description ◽

Quantitative Understanding ◽

Living Organisms ◽

Scientific Challenge

Gene regulation plays a central role in the development and functioning of living organisms. Developing a deeper qualitative and quantitative understanding of gene regulation is an important scientific challenge. The Lambda switch is commonly used as a paradigm of gene regulation. Verbal descriptions of the structure and functioning of the Lambda switch have appeared in biological textbooks. We apply fuzzy modeling to transform one such verbal description into a well-defined mathematical model. The resulting model is a piecewise-quadratic, second-order differential equation. It demonstrates functional fidelity with known results while being simple enough to allow a rather detailed analysis. Properties such as the number, location, and domain of attraction of equilibrium points can be studied analytically. Furthermore, the model provides a rigorous explanation for the so-called stability puzzle of the Lambda switch.

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The fimYZ Genes Regulate Salmonella enterica Serovar Typhimurium Invasion in Addition to Type 1 Fimbrial Expression and Bacterial Motility

Infection and Immunity ◽

10.1128/iai.73.3.1377-1385.2005 ◽

2005 ◽

Vol 73 (3) ◽

pp. 1377-1385 ◽

Cited By ~ 38

Author(s):

M. Aaron Baxter ◽

Bradley D. Jones

Keyword(s):

Gene Expression ◽

Gene Regulation ◽

Protein Interactions ◽

Salmonella Enterica ◽

Negative Regulator ◽

Invasive Phenotype ◽

Environmental Signals ◽

Bacterial Physiology ◽

Serovar Typhimurium

ABSTRACT An important step in Salmonella enterica serovar Typhimurium virulence is the ability to invade the intestinal epithelium. The invasion process requires a large number of genes encoded on Salmonella pathogenicity island 1 (SPI-1) at centisome 63 as well as genes located in other positions throughout the chromosome. Expression of the invasive phenotype is tightly regulated by environmental cues that are processed by a complex regulatory scheme. A central player in the invasion regulatory pathway is the HilA protein, which is transcriptional activator belonging to the OmpR/ToxR family. A number of positive regulators (hilC, hilD, fis, sirA/barA, csrAB, phoBR, fadD, envZ/ompR, and fliZ) and negative regulators (hha, hilE, lon, ams, phoP c and pag) have been identified that are able to alter expression of hilA transcription. Recent work has found that hilA transcription requires the HilD protein for activation. Other work has emphasized the importance of HilE as a negative regulator of hilA. Overexpression of hilE superrepresses hilA transcription, as well as the invasive phenotype. Two-hybrid experiments suggest that HilE exerts its regulatory influence on hilA through protein-protein interactions with HilD as the protein does not bind to the hilA promoter nor does it affect hilD transcription. As it seems likely that hilE plays an important role in translating environmental signals into invasion gene regulation, we have attempted to identify how the hilE gene itself is regulated. Our results indicate that the fimYZ genes, response regulatory proteins involved in type 1 fimbrial gene expression and recently implicated in motility gene regulation, are important activators of hilE expression. These findings indicate that invasion gene expression is coregulated with motility and adherence and provide experimental evidence that the expression of these virulence phenotypes is a subset of the overall regulation of bacterial physiology.

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