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Mapping the Tail Fiber as the Receptor Binding Protein Responsible for Differential Host Specificity of Pseudomonas aeruginosa Bacteriophages PaP1 and JG004

机译:映射尾纤维作为负责铜绿假单胞菌噬菌体PaP1和JG004的宿主特异性差异的受体结合蛋白。

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摘要

The first step in bacteriophage infection is recognition and binding to the host receptor, which is mediated by the phage receptor binding protein (RBP). Different RBPs can lead to differential host specificity. In many bacteriophages, such as Escherichia coli and Lactococcal phages, RBPs have been identified as the tail fiber or protruding baseplate proteins. However, the tail fiber-dependent host specificity in Pseudomonas aeruginosa phages has not been well studied. This study aimed to identify and investigate the binding specificity of the RBP of P. aeruginosa phages PaP1 and JG004. These two phages share high DNA sequence homology but exhibit different host specificities. A spontaneous mutant phage was isolated and exhibited broader host range compared with the parental phage JG004. Sequencing of its putative tail fiber and baseplate region indicated a single point mutation in ORF84 (a putative tail fiber gene), which resulted in the replacement of a positively charged lysine (K) by an uncharged asparagine (N). We further demonstrated that the replacement of the tail fiber gene (ORF69) of PaP1 with the corresponding gene from phage JG004 resulted in a recombinant phage that displayed altered host specificity. Our study revealed the tail fiber-dependent host specificity in P. aeruginosa phages and provided an effective tool for its alteration. These contributions may have potential value in phage therapy.
机译:噬菌体感染的第一步是识别并与宿主受体结合,这是由噬菌体受体结合蛋白(RBP)介导的。不同的RBP可能导致不同的宿主特异性。在许多噬菌体中,例如大肠埃希氏菌和乳球菌噬菌体,RBP被鉴定为尾纤维或突出的底板蛋白。然而,铜绿假单胞菌噬菌体中尾纤维依赖的宿主特异性尚未得到很好的研究。该研究旨在鉴定和研究铜绿假单胞菌噬菌体PaP1和JG004的RBP的结合特异性。这两个噬菌体具有很高的DNA序列同源性,但表现出不同的宿主特异性。与亲本噬菌体JG004相比,分离了自发突变噬菌体并显示出更广泛的宿主范围。推定的尾巴纤维和基板区域的测序表明ORF84(推定的尾巴纤维基因)发生单点突变,导致带正电荷的赖氨酸(K)被不带电荷的天冬酰胺(N)取代。我们进一步证明用来自噬菌体JG004的相应基因替换PaP1的尾巴纤维基因(ORF69)导致重组噬菌体表现出改变的宿主特异性。我们的研究揭示了铜绿假单胞菌噬菌体中尾纤维依赖的宿主特异性,并为其改变提供了有效的工具。这些贡献可能在噬菌体治疗中具有潜在价值。

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