Clostridioides difficile S-Layer Protein A (SlpA) Serves as a General Phage Receptor.
Clostridioides difficile S-Layer Protein A (SlpA) Serves as a General Phage Receptor.
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DOI：
10.1128/spectrum.03894-22
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发表时间：
2023-02-15
影响因子：
3.7
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中科院分区：
文献类型：
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治疗性噬菌体（噬菌体）被认为是对抗艰难梭菌感染的替代方法。为了提高噬菌体的效率，噬菌体必须具有广泛的宿主范围，但缺乏对艰难梭菌噬菌体使用的细胞受体的了解，阻碍了噬菌体鸡尾酒的合理设计。最近的报道表明艰难梭菌表面层蛋白A （SlpA）是一种重要的噬菌体受体，但可用的数据仍然有限。在这里，我们使用流行的R20291菌株及其缺乏功能s层的FM2.5突变衍生物，我们发现缺乏SlpA使细胞完全抵抗通常感染亲本菌株的<s:2> cd38 -2, <s:2> cd111和<s:2> cd146的感染。与从质粒中表达的12种不同的s层卡带型（slct）的互补表明，SLCT-6也允许被<s:3> cd111感染，而SLCT-11则允许被<s:3> cd38 -2和<s:3> cd146感染。值得注意的是，SLCT-1、-6、-8、-9、-10或-12的表达使5种通常不感染R20291菌株的噬菌体对感染产生易感性。此外，在SlpA的低分子量片段中，D2结构域的缺失被发现可以消除<s:2> cd38 -2和<s:2> cd146的感染，但不能消除<s:2> cd111的感染。总之，我们的数据表明，许多噬菌体使用SlpA作为它们的受体，最重要的是，尽管它们的尾部结构存在重大差异，但虹吸体和肌噬体都以SlpA为目标。因此，我们的研究代表了理解艰难梭菌与其噬菌体之间相互作用的重要一步。噬菌体治疗是治疗艰难梭菌感染的一种有趣的替代方法，因为与抗生素相反，大多数噬菌体具有高度的物种特异性，从而保护有益的肠道微生物免受感染。然而，目前可用的抗艰难梭菌噬菌体宿主范围狭窄，目标成员仅来自一种或几种PCR核糖型。如果不清楚确定宿主特异性的因素，特别是噬菌体识别的宿主受体，就很难以合理的方式开发治疗鸡尾酒。在我们的研究中，我们提供了清晰而明确的实验证据，证明SlpA是许多虹吸细胞和肌噬细胞共同使用的受体。尽管还需要确定特定噬菌体受体结合蛋白如何与特定的SLCT结合，但SlpA作为一种常见受体的鉴定是一个重要的关键，它将有助于合理设计针对临床重要菌株的治疗性噬菌体鸡尾酒。
Therapeutic bacteriophages (phages) are being considered as alternatives in the fight against Clostridioides difficile infections. To be efficient, phages should have a wide host range, buthe lack of knowledge about the cell receptor used by C. difficile phages hampers the rational design of phage cocktails. Recent reports suggested that the C. difficile surface layer protein A (SlpA) is an important phage receptor, but available data are still limited. Here, using the epidemic R20291 strain and its FM2.5 mutant derivative lacking a functional S-layer, we show that the absence of SlpA renders cells completely resistant to infection by ϕCD38-2, ϕCD111, and ϕCD146, which normally infect the parental strain. Complementation with 12 different S-layer cassette types (SLCTs) expressed from a plasmid revealed that SLCT-6 also allowed infection by ϕCD111 and SLCT-11 enabled infection by ϕCD38-2 and ϕCD146. Of note, the expression of SLCT-1, -6, -8, -9, -10, or -12 conferred susceptibility to infection by 5 myophages that normally do not infect the R20291 strain. Also, deletion of the D2 domain within the low-molecular-weight fragment of SlpA was found to abolish infection by ϕCD38-2 and ϕCD146 but not ϕCD111. Altogether, our data suggest that many phages use SlpA as their receptor and, most importantly, that both siphophages and myophages target SlpA despite major differences in their tail structures. Our study therefore represents an important step in understanding the interactions between C. difficile and its phages. IMPORTANCE Phage therapy represents an interesting alternative to treat Clostridioides difficile infections because, contrary to antibiotics, most phages are highly species specific, thereby sparing the beneficial gut microbes that protect from infection. However, currently available phages against C. difficile have a narrow host range and target members from only one or a few PCR ribotypes. Without a clear comprehension of the factors that define host specificity, and in particular the host receptor recognized by phages, it is hard to develop therapeutic cocktails in a rational manner. In our study, we provide clear and unambiguous experimental evidence that SlpA is a common receptor used by many siphophages and myophages. Although work is still needed to define how a particular phage receptor-binding protein binds to a specific SLCT, the identification of SlpA as a common receptor is a major keystone that will facilitate the rational design of therapeutic phage cocktails against clinically important strains.
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影响因子：
17.1
作者：
Kirk JA;Gebhart D;Buckley AM;Lok S;Scholl D;Douce GR;Govoni GR;Fagan RP
通讯作者：
Fagan RP
影响因子：
16.6
作者：
Lanzoni-Mangutchi P;Banerji O;Wilson J;Barwinska-Sendra A;Kirk JA;Vaz F;O'Beirne S;Baslé A;El Omari K;Wagner A;Fairweather NF;Douce GR;Bullough PA;Fagan RP;Salgado PS
通讯作者：
Salgado PS
影响因子：
4.8
作者：
Fagan, Robert P.;Fairweather, Neil F.
通讯作者：
Fairweather, Neil F.
影响因子：
6.4
作者：
Dingle, Kate E.;Didelot, Xavier;Crook, Derrick W.
通讯作者：
Crook, Derrick W.
影响因子：
3.2
作者：
Gebhart, Dana;Williams, Steven R.;Scholl, Dean
通讯作者：
Scholl, Dean