Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, OH, United States.
Infectious Diseases Institute, The Ohio State University, Columbus, OH, United States.
Front Cell Infect Microbiol. 2021 Jul 22;11:698146. doi: 10.3389/fcimb.2021.698146. eCollection 2021.
L-arabinose inducible promoters are commonly used in gene expression analysis. However, nutrient source and availability also play a role in biofilm formation; therefore, L-arabinose metabolism could impact biofilm development. In this study we examined the impact of L-arabinose on serovar Typhimurium (. Typhimurium) biofilm formation. Using mutants impaired for the transport and metabolism of L-arabinose, we showed that L-arabinose metabolism negatively impacts Typhimurium biofilm formation . When L-arabinose metabolism is abrogated, biofilm formation returned to baseline levels. However, without the ability to import extracellular L-arabinose, biofilm formation significantly increased. Using RNA-Seq we identified several gene families involved in these different phenotypes including curli expression, amino acid synthesis, and L-arabinose metabolism. Several individual candidate genes were tested for their involvement in the L-arabinose-mediated biofilm phenotypes, but most played no significant role. Interestingly, in the presence of L-arabinose the diguanylate cyclase gene A was downregulated in wild type . Typhimurium. Meanwhile , encoding an adenylate cyclase, was downregulated in an L-arabinose transport mutant. Using an IPTG-inducible plasmid to deplete c-di-GMP expression, we were able to abolish the increased biofilm phenotype seen in the transport mutant. However, the mechanism by which the L-arabinose import mutant forms significantly larger biofilms remains to be determined. Regardless, these data suggest that L-arabinose metabolism influences intracellular c-di-GMP levels and therefore biofilm formation. These findings are important when considering the use of an L-arabinose inducible promoter in biofilm conditions.
L-阿拉伯糖诱导启动子常用于基因表达分析。然而,营养物质的来源和可用性也在生物膜形成中起作用;因此,L-阿拉伯糖代谢可能会影响生物膜的发展。在本研究中,我们研究了 L-阿拉伯糖对鼠伤寒沙门氏菌(. Typhimurium)生物膜形成的影响。使用 L-阿拉伯糖转运和代谢缺陷的突变体,我们表明 L-阿拉伯糖代谢对鼠伤寒沙门氏菌生物膜形成有负面影响。当 L-阿拉伯糖代谢被阻断时,生物膜形成恢复到基线水平。然而,没有能力导入细胞外的 L-阿拉伯糖,生物膜形成显著增加。使用 RNA-Seq,我们确定了几个参与这些不同表型的基因家族,包括卷曲表达、氨基酸合成和 L-阿拉伯糖代谢。测试了几个候选基因在 L-阿拉伯糖介导的生物膜表型中的作用,但大多数没有发挥重要作用。有趣的是,在存在 L-阿拉伯糖的情况下,野生型鼠伤寒沙门氏菌中的二鸟苷酸环化酶基因 A 被下调。与此同时,编码腺苷酸环化酶的基因在 L-阿拉伯糖转运突变体中被下调。使用 IPTG 诱导的质粒耗尽 c-di-GMP 表达,我们能够消除在转运突变体中观察到的增加的生物膜表型。然而,L-阿拉伯糖进口突变体形成更大生物膜的机制仍有待确定。无论如何,这些数据表明 L-阿拉伯糖代谢影响细胞内 c-di-GMP 水平,从而影响生物膜形成。在考虑在生物膜条件下使用 L-阿拉伯糖诱导启动子时,这些发现很重要。