Colanic acid and lipopolysaccharide in Pectobacterium carotovorum Pcc21 serve as receptors for the bacteriophage phiPccP-2

IF 6.1 1区生物学 Q1 MICROBIOLOGY

Microbiological research Pub Date : 2024-10-28 DOI:10.1016/j.micres.2024.127939

Nguyen Trung Vu , Hyeongsoon Kim , In Sun Hwang , Chang-Sik Oh

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Abstract

Bacteriophages (phages) are viruses that specifically bind to and infect target bacteria. The phage phiPccP-2, belonging to the Myoviridae family, efficiently controls Pectobacterium spp. In the present study, we aimed to elucidate the mechanism of recognition of P. carotovorum Pcc21 by phiPccP-2. The EZ-Tn5 transposon mutant library of Pcc21 was used to screen for phage-resistant mutants. Among 4072 mutants screened, 12 harbored disruptions in genes associated with the biosynthesis of either colanic acid (CA) or lipopolysaccharide (LPS) showed resistance to phiPccP-2. Complementation of 4 representative phage-resistant mutants with the corresponding genes fully restored the binding ability and lytic activity of PhiPccP-2. The amounts of CA or LPS structure in these mutants were significantly altered compared with those in the wild-type strain. Adsorption competition assays between CA and LPS extracted from Pcc21 and the natural receptors in Pcc21 showed that unbound phages were significantly increased, indicating that both CA and LPS are associated with the adsorption of the phiPccP-2 to Pcc21. In contrast, the adsorption of phiPccP-2 to extracted CA or LPS did not inactivate the lytic activity of phiPccP-2, indicating that the adsorption to the extracted CA or LPS is not sufficient for DNA injection. Treatment with polymyxin B, which disrupts LPS, interfered with phiPccP-2 adsorption to Pcc21. Furthermore, phage-resistant mutants showed reduced virulence in the host plant, suggesting a trade-off between phage resistance and bacterial virulence. Overall, our results indicate that both CA and LPS serve as receptors for the binding of phiPccP-2 to P. carotovorum Pcc21.

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果胶杆菌 Pcc21 中的胶酸和脂多糖是噬菌体 phiPccP-2 的受体。

噬菌体（噬菌体）是一种能特异性结合并感染目标细菌的病毒。本研究旨在阐明 phiPccP-2 识别 P. carotovorum Pcc21 的机制。我们利用 Pcc21 的 EZ-Tn5 转座子突变体文库筛选噬菌体抗性突变体。在筛选出的 4072 个突变体中，有 12 个突变体与可乐酸（CA）或脂多糖（LPS）的生物合成有关，它们对 phiPccP-2 具有抗性。用相应的基因对 4 个具有代表性的噬菌体抗性突变体进行补体，完全恢复了 PhiPccP-2 的结合能力和溶解活性。与野生型菌株相比，这些突变体中的 CA 或 LPS 结构量发生了显著变化。从 Pcc21 中提取的 CA 和 LPS 与 Pcc21 中的天然受体之间的吸附竞争试验表明，未结合的噬菌体明显增加，这表明 CA 和 LPS 都与 phiPccP-2 对 Pcc21 的吸附有关。相反，phiPccP-2吸附在提取的CA或LPS上并不会使phiPccP-2的溶菌活性失活，这表明吸附在提取的CA或LPS上不足以进行DNA注入。用能破坏 LPS 的多粘菌素 B 处理也会干扰 phiPccP-2 对 Pcc21 的吸附。此外，噬菌体抗性突变体在寄主植物中的毒力也有所降低，这表明噬菌体抗性与细菌毒力之间存在权衡。总之，我们的研究结果表明，CA 和 LPS 都是 phiPccP-2 与 P. carotovorum Pcc21 结合的受体。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microbiological research 生物-微生物学

CiteScore

10.90

自引率

6.00%

发文量

249

审稿时长

29 days

期刊介绍： Microbiological Research is devoted to publishing reports on prokaryotic and eukaryotic microorganisms such as yeasts, fungi, bacteria, archaea, and protozoa. Research on interactions between pathogenic microorganisms and their environment or hosts are also covered.