铜绿假单胞菌中假定的 NCS1 和 NCS2 核碱基转运体的底物鉴定。

IF 5.1 1区 生物学 Q1 MICROBIOLOGY mBio Pub Date : 2024-10-30 DOI:10.1128/mbio.02434-24
Corey Kennelly, Arthur Prindle
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引用次数: 0

摘要

铜绿假单胞菌是一种机会性病原体,它可以从环境中回收核碱基,以节省原本用于新核苷酸生物合成的营养物质。然而,人们对铜绿微囊藻中 13 种推测的核碱基转运体的底物特异性知之甚少。在这里,我们结合使用遗传和化学方法,报告了铜绿假单胞菌中 10 种推定核碱基转运体的底物鉴定结果。具体来说,我们在缺乏所有 13 个推定核碱基转运体的遗传背景中分别表达了每个转运体,并对以 10 种核碱基为唯一氮源的生长进行了量化。我们利用靶向基因敲除确认了这些基于表达的底物鉴定。作为补充,我们利用四种有毒的核碱基抗代谢物来鉴定这些菌株的抗菌活性。我们发现了唯一的尿囊素转运体以及鸟嘌呤、黄嘌呤、尿酸、胞嘧啶、胸腺嘧啶、尿嘧啶和二氢尿嘧啶的转运体。此外,我们还发现至少有五个核碱基转运体与次黄嘌呤有关,最近有报道称次黄嘌呤是铜绿假单胞菌的一种抗生物膜线索。这些结果提供了铜绿假单胞菌中假定核碱基转运体的初步特征,极大地推动了我们对这种临床相关生物体中核碱基转运的了解:重要意义:铜绿假单胞菌是一种经常具有多重耐药性的机会性病原体,也是最常见的医源性感染病因之一。众所周知,核碱基可在营养有限的条件下支持生长,但最近的研究表明,腺嘌呤和次黄嘌呤也可通过破坏 c-di-GMP 代谢来减少铜绿假单胞菌生物膜的形成。因此,核碱基转运可能与铜绿假单胞菌生物学和致病机理的多个方面有关。然而,目前人们对铜绿假单胞菌体内核碱基的转运知之甚少。我们的研究报告初步鉴定了铜绿假单胞菌中 10 个假定的核碱基转运体的底物,为解决以前难以检验的有关该生物体核碱基转运的假说提供了新的工具。
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Substrate identification of putative NCS1 and NCS2 nucleobase transporters in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic pathogen that can salvage nucleobases from the environment to conserve nutrients that would otherwise be spent on de novo nucleotide biosynthesis. However, little is known regarding the substrate specificity of the 13 putative nucleobase transporters in P. aeruginosa. Here, using a combination of genetic and chemical approaches, we report substrate identifications for 10 putative nucleobase transporters in P. aeruginosa. Specifically, we individually expressed each transporter in a genetic background lacking all 13 putative nucleobase transporters and quantified growth on a panel of 10 nucleobases as sole nitrogen sources. We confirmed these expression-based substrate identifications using targeted genetic knockouts. In a complementary approach, we utilized four toxic nucleobase antimetabolites to characterize antimicrobial activity in these same strains. We identified the sole allantoin transporter as well as transporters for guanine, xanthine, uric acid, cytosine, thymine, uracil, and dihydrouracil. Furthermore, we associated at least five nucleobase transporters with hypoxanthine, which has been recently reported to be an antibiofilm cue in P. aeruginosa. These results provide an initial characterization of the putative nucleobase transporters in P. aeruginosa, significantly advancing our understanding of nucleobase transport in this clinically relevant organism.

Importance: Pseudomonas aeruginosa is a frequently multidrug-resistant opportunistic pathogen and one of the most common causes of healthcare-acquired infections. While nucleobases are known to support growth in nutrient-limited conditions, recent work showed that adenine and hypoxanthine can also decrease P. aeruginosa biofilm formation by disrupting c-di-GMP metabolism. Thus, nucleobase transport may be relevant to multiple aspects of P. aeruginosa biology and pathogenesis. However, there is currently little known about the transport of nucleobases in P. aeruginosa. Our work reports initial substrate identifications for 10 putative nucleobase transporters in P. aeruginosa, providing new tools to address previously difficult-to-test hypotheses relating to nucleobase transport in this organism.

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来源期刊
mBio
mBio MICROBIOLOGY-
CiteScore
10.50
自引率
3.10%
发文量
762
审稿时长
1 months
期刊介绍: mBio® is ASM''s first broad-scope, online-only, open access journal. mBio offers streamlined review and publication of the best research in microbiology and allied fields.
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