微氧条件下Dnr调控的反硝化作用。

Dnr-regulated denitrification in microoxic conditions.

作者信息

Stuut Balsam Stacie, Conaway Amy, Mould Dallas L, Jean-Pierre Fabrice, Hogan Deborah A

机构信息

Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA.

出版信息

Microbiol Spectr. 2025 Sep 2;13(9):e0068225. doi: 10.1128/spectrum.00682-25. Epub 2025 Aug 7.

Abstract

causes acute and chronic infections, such as those that occur in the lungs of people with cystic fibrosis (CF). In infection environments, oxygen (O) concentrations are often low. The transcription factor Anr (naerobic regulation of arginine deiminase and itrate eduction) responds to low O by upregulating genes necessary for fitness in microoxic and anoxic conditions. Anr regulates Dnr (issimilative itrate respiration egulator), a gene encoding a transcriptional regulator that promotes the expression of genes required for using nitrate as an alternative electron acceptor during denitrification. In CF sputum, transcripts involved in denitrification are highly expressed. While Dnr is necessary for the anoxic growth of in CF sputum and artificial sputum media (ASMi), the contribution of denitrification to fitness in oxic conditions has not been well described. Here, we show that requires for fitness in ASMi, and the requirement for is abolished when nitrate is excluded from the media. Additionally, we show that consumes nitrate in lysogeny broth (LB) under microoxic conditions. Furthermore, strains without a functioning quorum sensing regulator LasR, which leads to elevated Anr activity, consume nitrate in LB even in normoxia. There was no growth advantage for when nitrate was present at concentrations from 100 to 1,600 µM. However, consumption of nitrate in oxic conditions created a requirement for Dnr and Dnr-regulated NorCB, likely due to the need to detoxify nitric oxide. These studies suggest that Anr- and Dnr-regulated processes may impact physiology in many common culture conditions.IMPORTANCE is an opportunistic pathogen commonly isolated from low-oxygen environments such as the lungs of people with cystic fibrosis. While the importance of energy generation by denitrification is clear in anoxic environments, the effects of denitrification in oxic cultures are not well understood. Here, we show that nitrate is consumed in microoxic environments and, in some strains, in normoxic environments. While nitrate does not appear to stimulate microoxic growth rate or yield, it does impact physiology. We show that the regulators Anr (naerobic regulation of arginine deiminase and itrate eduction) and Dnr (issimilative itrate respiration egulator), which are best known for their roles in anoxic conditions, contribute to fitness in common laboratory media in the presence of oxygen.

摘要

可引发急性和慢性感染,比如在患有囊性纤维化(CF)的人的肺部发生的感染。在感染环境中,氧气(O₂)浓度通常较低。转录因子Anr(精氨酸脱亚氨酶和硝酸盐还原的厌氧调节)通过上调在微氧和缺氧条件下生存所需的基因来响应低O₂水平。Anr调节Dnr(异化硝酸盐呼吸调节因子),Dnr是一个编码转录调节因子的基因,该转录调节因子可促进在反硝化过程中使用硝酸盐作为替代电子受体所需基因的表达。在CF痰液中,参与反硝化作用的转录本高度表达。虽然Dnr对于在CF痰液和人工痰液培养基(ASMi)中缺氧生长是必需的,但反硝化作用对在有氧条件下生存能力的贡献尚未得到充分描述。在此,我们表明在ASMi中生存需要Dnr,并且当培养基中不含硝酸盐时,对Dnr的需求就会消失。此外,我们表明在微氧条件下,[细菌名称]在溶菌肉汤(LB)中会消耗硝酸盐。此外,缺乏功能性群体感应调节因子LasR(这会导致Anr活性升高)的菌株,即使在常氧条件下也会在LB中消耗硝酸盐。当硝酸盐浓度在100至1600µM之间时,[细菌名称]没有生长优势。然而,在有氧条件下[细菌名称]消耗硝酸盐产生了对Dnr和Dnr调节的NorCB的需求,这可能是由于需要对一氧化氮进行解毒。这些研究表明,Anr和Dnr调节的过程可能会在许多常见培养条件下影响[细菌名称]的生理状态。重要性[细菌名称]是一种机会性病原体,通常从低氧环境中分离出来,如患有囊性纤维化的人的肺部。虽然在缺氧环境中通过反硝化作用产生能量的重要性是明确的,但反硝化作用在有氧培养中的影响尚未得到充分理解。在此,我们表明在微氧环境中以及在某些菌株的常氧环境中会消耗硝酸盐。虽然硝酸盐似乎不会刺激微氧生长速率或产量,但它确实会影响生理状态。我们表明,以在缺氧条件下的作用最为人所知的调节因子Anr(精氨酸脱亚氨酶和硝酸盐还原的厌氧调节)和Dnr(异化硝酸盐呼吸调节因子),在有氧存在的情况下对[细菌名称]在常见实验室培养基中的生存能力有贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6781/12403760/83548312b738/spectrum.00682-25.f001.jpg

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