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存在于有益细菌中的有害噬菌体:神秘的λ噬菌体和志贺毒素转化噬菌体Φ24的作用

Bad Phages in Good Bacteria: Role of the Mysterious of λ and Shiga Toxin-Converting Φ24 Bacteriophages.

作者信息

Dydecka Aleksandra, Bloch Sylwia, Rizvi Ali, Perez Shaili, Nejman-Falenczyk Bozena, Topka Gracja, Gasior Tomasz, Necel Agnieszka, Wegrzyn Grzegorz, Donaldson Logan W, Wegrzyn Alicja

机构信息

Department of Molecular Biology, Faculty of Biology, University of GdanskGdansk, Poland.

Department of Biology, York UniversityToronto, ON, Canada.

出版信息

Front Microbiol. 2017 Aug 25;8:1618. doi: 10.3389/fmicb.2017.01618. eCollection 2017.

Abstract

Lambdoid bacteriophages form a group of viruses that shares a common schema of genome organization and lifecycle. Some of them can play crucial roles in creating the pathogenic profiles of strains. For example, Shiga toxin-producing (STEC) acquired genes, encoding Shiga toxins, via lambdoid prophages (Stx phages). The results obtained so far present the evidence for the relation between the region of the phage genome and lambdoid phage development, however molecular mechanisms of activities of the genes' products are still unknown. In view of this, we decided to determine the influence of the uncharacterized open reading frame of the region on lambdoid phages development using recombinant prophages, λ and Stx phage Φ24 We have demonstrated that codes for a folded protein, thus, it is a functional gene. NMR spectroscopy and analytical gel filtration were used to extend this observation further. From backbone chemical shifts, Orf63 is oligomeric in solution, likely a trimer and consistent with its small size (63 aa.), is comprised of two helices, likely intertwined to form the oligomer. We observed that the deletion of phage does not impair the intracellular lambdoid phage lytic development, however delays the time and decreases the efficiency of prophage induction and in consequence results in increased survival of during phage lytic development. Additionally, the deletion of phage negatively influences expression of the major phage genes and open reading frames from the region during prophage induction with hydrogen peroxide. We conclude, that lambdoid phage may have specific functions in the regulation of lambdoid phages development, especially at the stage of the lysis vs. lysogenization decision. Besides, probably participates in the regulation of the level of expression of essential phage genes and open reading frames from the region during prophage induction.

摘要

λ噬菌体形成了一组具有共同基因组组织模式和生命周期的病毒。其中一些在塑造菌株的致病特征方面可发挥关键作用。例如,产志贺毒素大肠杆菌(STEC)通过λ样原噬菌体(Stx噬菌体)获得了编码志贺毒素的基因。目前获得的结果为噬菌体基因组的某个区域与λ样噬菌体发育之间的关系提供了证据,然而该区域基因产物的分子作用机制仍不清楚。有鉴于此,我们决定使用重组原噬菌体λ和Stx噬菌体Φ24来确定该区域未表征的开放阅读框对λ样噬菌体发育的影响。我们已经证明该开放阅读框编码一种折叠蛋白,因此它是一个功能基因。利用核磁共振光谱和分析凝胶过滤进一步扩展了这一观察结果。从主链化学位移来看,Orf63在溶液中是寡聚体,可能是三聚体,并且鉴于其较小的尺寸(63个氨基酸),由两个螺旋组成,可能相互缠绕形成寡聚体。我们观察到噬菌体该区域的缺失并不损害细胞内λ样噬菌体的裂解发育,但会延迟时间并降低原噬菌体诱导的效率,从而导致在噬菌体裂解发育过程中该区域的存活率增加。此外,在用过氧化氢诱导原噬菌体时,噬菌体该区域的缺失对主要噬菌体基因和该区域开放阅读框的表达有负面影响。我们得出结论,λ样噬菌体该区域可能在调节λ样噬菌体发育方面具有特定功能,特别是在裂解与溶原化决定阶段。此外,该区域可能在原噬菌体诱导期间参与调节必需噬菌体基因和该区域开放阅读框的表达水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/74e2/5575149/9b41bdf39606/fmicb-08-01618-g0001.jpg

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