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Phosphotransferase System-Dependent Extracellular Growth of Listeria monocytogenes Is Regulated by Alternative Sigma Factors σL and σH

机译:依赖于磷酸转移酶系统的单核细胞增生李斯特菌的细胞外生长受其他Sigma因子σL和σH调节

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Alternative sigma (σ) factors and phosphotransferase systems (PTSs) play pivotal roles in the environmental adaptation and virulence of Listeria monocytogenes . The growth of the L. monocytogenes parent strain 10403S and 15 isogenic alternative σ factor mutants was assessed in defined minimal medium (DM) with PTS-dependent or non-PTS-dependent carbon sources at 25°C or 37°C. Overall, our results suggested that the regulatory effect of alternative σ factors on the growth of L. monocytogenes is dependent on the temperature and the carbon source. One-way analysis of variance (one-way ANOVA) showed that the factor “strain” had a significant effect on the maximum growth rate (μ_(max)), lag phase duration (λ), and maximum optical density (OD_(max)) in PTS-dependent carbon sources ( P < 0.05) but not in a non-PTS-dependent carbon source. Also, the OD_(max) was not affected by strain for any of the three PTS-dependent carbon sources at 25°C but was affected by strain at 37°C. Monitoring by quantitative real-time PCR (qRT-PCR) showed that transcript levels for lmo0027 , a glucose-glucoside PTS permease (PTS~(Glc)-1)-encoding gene, were higher in the absence of σ~(L), and lower in the absence of σ~(H), than in the parent strain. Our data thus indicate that σ~(L) negatively regulates lmo0027 and that the increased μ_(max) observed for the Δ sigL strain in DM with glucose may be associated with increased expression of PTS~(Glc)-1 encoded by lmo0027 . Our findings suggest that σ~(H) and σ~(L) mediate the PTS-dependent growth of L. monocytogenes through complex transcriptional regulations and fine-tuning of the expression of specific pts genes, including lmo0027 . Our findings also reveal a more important and complex role of alternative σ factors in the regulation of growth in different sugar sources than previously assumed.
机译:选择性σ(σ)因子和磷酸转移酶系统(PTS)在单核细胞增多性李斯特菌的环境适应和毒性中起着关键作用。在25°C或37°C下,在具有PTS依赖性或非PTS依赖性碳源的限定基本培养基(DM)中,评估了单核细胞增生李斯特氏菌亲本菌株10403S和15个等基因替代σ因子突变体的生长。总体而言,我们的结果表明,替代性σ因子对单核细胞增生李斯特菌生长的调节作用取决于温度和碳源。单向方差分析(单向方差分析)表明,“应变”因子对最大生长速率(μ_(max)),滞后相持续时间(λ)和最大光密度(OD_(max ))在PTS依赖的碳源中(P <0.05),但在非PTS依赖的碳源中则没有。另外,对于25°C时三种依赖PTS的碳源,OD_(max)不受应变的影响,但受37°C的应变的影响。实时定量PCR(qRT-PCR)监测显示,在没有σ〜(L)的情况下,lmo0027(一种葡萄糖-葡萄糖苷PTS渗透酶(PTS〜(Glc)-1)编码基因)的转录水平更高,在没有σ〜(H)的情况下比在亲本菌株中要低。因此,我们的数据表明,σ〜(L)负调控lmo0027,并且在DM中用葡萄糖观察到的ΔsigL菌株增加的μ_(max)可能与lmo0027编码的PTS〜(Glc)-1的表达增加有关。我们的发现表明,σ〜(H)和σ〜(L)通过复杂的转录调控和微调特定pts基因(包括lmo0027)的表达来介导单核细胞增生李斯特菌的PTS依赖性生长。我们的发现还揭示了替代性σ因子在调节不同糖源中的生长方面比以前假设的更为重要和复杂的作用。

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