PA5339, a RidA Homolog, Is Required for Full Growth in Pseudomonas aeruginosa
PA5339, a RidA Homolog, Is Required for Full Growth in Pseudomonas aeruginosa
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DOI：
10.1128/jb.00434-18
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发表时间：
2018-11-01
影响因子：
3.2
通讯作者：
Downs, Diana M.
中科院分区：
文献类型：
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作者：
Irons, Jessica;Hodge-Hanson, Kelsey M.;Downs, Diana M.
Rid蛋白超家族（YJgF/YER 057 c/UK 114）存在于生命的所有领域。原型蛋白质是来自肠道沙门氏菌的RidA，是一种脱氨酶，可以猝灭反应性代谢物2-氨基丙烯酸酯（2AA）。2AA脱氨酶活性在来自人、植物、酵母、古细菌和细菌的RidA蛋白中是保守的。缺乏功能性RidA的肠道沙门氏菌、大肠杆菌和酿酒酵母的突变体表现出生长缺陷，表明2AA代谢应激同样保守。PubSEED数据库显示铜绿假单胞菌（PAO 1）编码Rid超家族的8个成员。筛选缺乏五种Rid蛋白中的每一种的铜绿假单胞菌PAO 1的突变体，并解剖在不存在PA 5339的情况下出现的突变体表型。PA 5339：：Tn突变体具有生长、运动和生物膜缺陷，这些缺陷都可能与2AA的积累有关。此外，PA 5339蛋白在体外被证明是2AA脱氨酶，并恢复代谢平衡为S。肠道病毒ridA突变体的体内研究。这里提供的数据表明，铜绿假单胞菌中的RidA范式与其他生物体中描述的相似，但不同之处在于仅删除多个同源物中的一个产生缺陷。重要信息RidA是一种广泛保守的蛋白质，可防止原核生物和真核生物中2-氨基丙烯酸酯（2AA）损伤吡哆醛5 '-磷酸（PLP）依赖性酶引起的内源性代谢应激。用于理解2AA的积累及其后果的框架在很大程度上已经在肠道沙门氏菌中定义。我们在这里表明，在铜绿假单胞菌（PAO 1），2AA积累导致生长减少，受损的运动性，和有缺陷的生物膜形成。这项研究扩展了我们的知识，生物体的代谢结构如何有助于PLP依赖性酶的2AA失活的后果，并确定了铜绿假单胞菌中的关键RidA蛋白。
The Rid protein superfamily (YJgF/YER057c/UK114) is found in all domains of life. The archetypal protein, RidA from Salmonella enterica, is a deaminase that quenches the reactive metabolite 2-aminoacrylate (2AA). 2AA deaminase activity is conserved in RidA proteins from humans, plants, yeast, archaea, and bacteria. Mutants of Salmonella enterica, Escherichia coli, and Saccharomyces cerevisiae that lack a functional RidA exhibit growth defects, suggesting that 2AA metabolic stress is similarly conserved. The PubSEED database shows Pseudomonas aeruginosa (PAO1) encodes eight members of the Rid superfamily. Mutants of P. aeruginosa PAO1 lacking each of five Rid proteins were screened, and the mutant phenotypes that arose in the absence of PA5339 were dissected. A PA5339::Tn mutant has growth, motility, and biofilm defects that can all be linked to the accumulation of 2AA. Further, the PA5339 protein was demonstrably a 2AA deaminase in vitro and restored metabolic balance to a S. enterica ridA mutant in vivo. The data presented here show that the RidA paradigm in Pseudomonas aeruginosa had similarities to those described in other organisms but was distinct in that deleting only one of multiple homologs generated deficiencies. Based on the collective data presented here in, PA5339 was renamed RidA.IMPORTANCE RidA is a widely conserved protein that prevents endogenous metabolic stress caused by 2-aminoacrylate (2AA) damage to pyridoxal 5'-phosphate (PLP)-dependent enzymes in prokaryotes and eukaryotes. The framework for understanding the accumulation of 2AA and its consequences have largely been defined in Salmonella enterica. We show here that in P. aeruginosa (PAO1), 2AA accumulation leads to reduced growth, compromised motility, and defective biofilm formation. This study expands our knowledge how the metabolic architecture of an organism contributes to the consequences of 2AA inactivation of PLP-dependent enzymes and identifies a key RidA protein in P. aeruginosa.