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Structural characterization of T7 tail machinery reveals a conserved tubular structure among other Podoviridae family members and suggests a common mechanism for DNA delivery T7尾部机械的结构特征揭示了Podoviridae家族成员中保守的管状结构,并提示了DNA传递的共同机制
Pub Date : 2013-10-01 DOI: 10.4161/bact.27011
A. Cuervo, M. Chagoyen, Mar Pulido-Cid, A. Camacho, J. Carrascosa
Bacteriophage tail complexes play an essential role in host recognition and DNA delivery during virus infection. These molecular machines are composed of a tubular structure surrounded by fibers, with a central channel that acts as a conduit for DNA ejection. The T7 tail complex is formed by four proteins: connector (gp8), gatekeeper (gp11), nozzle (gp12), and fibers (gp17). Previous biochemical and structural studies allowed definition of the stoichiometry and order of assembly of these proteins. Here we compared the tail complex from other Podoviridae phages that infect bacteria with Gram− type envelopes (K1E, P-SSP7, and ε15), and found strong similarities with the T7 nozzle; this was supported by sequence alignment and secondary structure prediction studies. These similarities were not observed in the new reconstruction of protein p9 presented here, which builds the hexameric nozzle of ϕ29, a virus that infects Gram+ bacteria. The results suggest that the Podoviridae nozzle has evolved to adapt to membrane composition of the infected host.
在病毒感染过程中,噬菌体尾部复合物在宿主识别和DNA传递中起着重要作用。这些分子机器是由一个被纤维包围的管状结构组成的,有一个中央通道作为DNA喷射的管道。T7尾部复合体由四种蛋白质组成:连接器(gp8)、看门人(gp11)、喷嘴(gp12)和纤维(gp17)。先前的生化和结构研究允许对这些蛋白质的化学计量和组装顺序进行定义。本文比较了其他具有革兰氏型包膜(K1E、P-SSP7和ε15)的足病毒科噬菌体的尾部复合体,发现与T7喷嘴有很强的相似性;这得到了序列比对和二级结构预测研究的支持。在构建感染革兰氏阳性细菌的病毒- - - 29的六聚体喷嘴的蛋白质p9的新重建中没有观察到这些相似性。结果表明,足病毒科的喷嘴已经进化到适应被感染宿主的膜组成。
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引用次数: 5
Life in Science: Björn H Lindqvist. 科学生活:Björn H Lindqvist。
Pub Date : 2013-10-01 DOI: 10.4161/bact.26673
Björn H Lindqvist
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引用次数: 0
Bacteriophage K for reduction of Staphylococcus aureusbiofilm on central venous catheter material. 噬菌体K用于减少中心静脉导管材料上的金黄色葡萄球菌生物膜。
Pub Date : 2013-10-01 Epub Date: 2013-10-24 DOI: 10.4161/bact.26825
Matthew P Lungren, Diana Christensen, Ravi Kankotia, Irene Falk, Ben E Paxton, Charles Y Kim

The purpose of this project was to determine whether bacteriophage can reduce bacterial colonization and biofilm formation on central venous catheter material. Twenty silicone discs were inoculated for 24 h with broth culture of Methicillin sensitive staphylococcus aureus (0.5 McFarland standard). The inoculate was aspirated and discs placed into two equal groups for 24 h: (1) untreated controls; (2) bacteriophage treatment (staphylococcal bacteriophage K, propagated titer > 108). At the completion of the experiment discs were processed for quantitative culture. Statistical testing was performed using the rank sum test. Mean colony forming units (CFU) were significantly decreased in experimental compared with controls (control 6.3 × 105 CFU, experimental 6.7 × 101, P ≤ 0.0001). Application of bacteriophage to biofilm infected central venous catheter material significantly reduced bacterial colonization and biofilm presence. Our data suggests that bacteriophage treatment may be a feasible strategy for addressing central venous catheter staph aureus biofilm infections.

本项目的目的是确定噬菌体是否可以减少中心静脉导管材料上细菌的定植和生物膜的形成。用甲氧西林敏感金黄色葡萄球菌(0.5 McFarland标准)肉汤培养物接种20个硅胶片24 h。抽吸接种物,将接种盘分成两组,放置24 h:(1)对照组;(2)噬菌体处理(葡萄球菌噬菌体K,繁殖滴度> 108)。实验完成后,对培养皿进行定量培养。采用秩和检验进行统计检验。试验组平均菌落形成单位(CFU)显著低于对照组(对照组6.3 × 105 CFU,试验组6.7 × 101, P≤0.0001)。将噬菌体应用于生物膜感染的中心静脉导管材料可显著减少细菌定植和生物膜的存在。我们的数据表明,噬菌体治疗可能是解决中心静脉导管金黄色葡萄球菌生物膜感染的可行策略。
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引用次数: 27
Cyanophages infecting Anabaena circinalis and Anabaena cylindrica in a tropical reservoir 热带水库中感染环状水蛭和圆柱水蛭的噬藻体
Pub Date : 2013-07-11 DOI: 10.4161/bact.25571
B. Yeo, K. Gin
A total of six lytic cyanophages have been isolated from Kranji Reservoir in Singapore. Two cyanophages infected a toxic species of cyanobacterium, A. circinalis, while four cyanophages infected the cyanobacterium A. cylindrica. To obtain the cyanophages that infect A. circinalis and A. cylindrica, two isolation methods were established. Cyanophages infecting A. circinalis were successfully isolated with well assay, and cyaophages infecting A. cylindrica were successfully isolated with double layer plaque assay. These isolation methods resulted in a rapid and significant lytic effect. A. circinalis cyanophage isolates are potential biocontrol of A. circinalis, due to the ability of cyanophage isolates in suppressing the growth of A. circinalis effectively. During the isolation process, we observed that A. cylindrica experienced changes in aggregation and algal mat formation when inoculated with the cyanophage isolates.
从新加坡Kranji水库共分离出6个溶藻体。两个噬藻体感染了一种有毒的蓝藻——环状藻,而四个噬藻体感染了白茅蓝藻。为获得侵染环状假单胞菌和圆柱假单胞菌的噬藻菌,建立了两种分离方法。用孔法成功分离了侵染线虫的噬菌体,用双层菌斑法成功分离了侵染线虫的噬菌体。这些分离方法具有快速、显著的裂解效果。环状吸青体分离物可有效抑制环状吸青体的生长,具有潜在的生物防治潜力。在分离过程中,我们观察到,当与噬藻菌分离物接种时,圆柱孢杆菌的聚集和藻席形成发生了变化。
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引用次数: 9
Tale of the unseen phage 看不见的噬菌体的故事
Pub Date : 2013-07-11 DOI: 10.4161/bact.25985
J. Mahony, S. Ainsworth, D. van Sinderen
Lactococcal phages are among the best characterized phages of Gram-positive bacteria and as such represent excellent models to promote our understanding of phage-host interactions. In recent years, microarray technology has been employed to define the transcriptional profile of phages during infection, while simultaneously uncovering the response of the host to the infection event. The responses to infection by a lytic and a temperate phage of Lactococcus lactis were analyzed, combined with an assessment of the host response to lysogenisation with the temperate phage, Tuc2009. Here, we discuss the lessons learned from such transcriptional profiling studies, focusing on studies relating to phages of Lactococcus lactis, and how this knowledge may be applied to future studies.
乳球菌噬菌体是革兰氏阳性菌中最具特征的噬菌体之一,因此代表了促进我们对噬菌体-宿主相互作用的理解的优秀模型。近年来,微阵列技术已被用于确定噬菌体在感染过程中的转录谱,同时揭示宿主对感染事件的反应。分析了乳酸乳球菌裂解噬菌体和温和噬菌体对感染的反应,并评估了宿主对温和噬菌体溶原作用的反应,Tuc2009。在这里,我们讨论了从这些转录谱研究中吸取的教训,重点是与乳酸乳球菌噬菌体有关的研究,以及如何将这些知识应用于未来的研究。
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引用次数: 1
Bacteriophages lytic for Salmonella rapidly reduce Salmonella contamination on glass and stainless steel surfaces. 沙门氏菌噬菌体可迅速减少玻璃和不锈钢表面的沙门氏菌污染。
Pub Date : 2013-07-01 Epub Date: 2013-07-26 DOI: 10.4161/bact.25697
Joelle Woolston, Adam R Parks, Tamar Abuladze, Bradley Anderson, Manrong Li, Chandi Carter, Leigh Farris Hanna, Serena Heyse, Duane Charbonneau, Alexander Sulakvelidze
A cocktail of six lytic bacteriophages, SalmoFresh™, significantly (p < 0.05) reduced the number of surface-applied Salmonella Kentucky and Brandenburg from stainless steel and glass surfaces by > 99% (2.1–4.3 log). Both strains were susceptible to SalmoFresh™ in the spot-test assay. Conversely, SalmoFresh™ was unable to reduce surface contamination with a Salmonella Paratyphi B strain that was not susceptible to the phage cocktail in the spot-test assay. However, by replacing two SalmoFresh™ component phages with two new phages capable of lysing the Paratyphi B strain in the spot-test assay, the target range of the cocktail was shifted to include the Salmonella Paratyphi B strain. The modified cocktail, SalmoLyse™, was able to significantly (p < 0.05) reduce surface contamination of the Paratyphi B strain by > 99% (2.1–4.1 log). The data show that both phage cocktails were effective in significantly reducing the levels of Salmonella on hard surfaces, provided the contaminating strains were susceptible in the spot-test (i.e., spot-test susceptibility was indicative of efficacy in subsequent surface decontamination studies). The data also support the concept that phage preparations can be customized to meet the desired antibacterial application.
六种溶解噬菌体SalmoFresh™的混合物显著(p < 0.05)减少了表面应用于不锈钢和玻璃表面的肯塔基沙门氏菌和勃兰登堡沙门氏菌的数量> 99% (2.1-4.3 log)。在现场试验中,两株菌株对SalmoFresh™敏感。相反,SalmoFresh™无法减少对噬菌体混合物不敏感的副伤寒沙门氏菌B株的表面污染。然而,通过在现场试验中将两个SalmoFresh™成分噬菌体替换为两个能够裂解副伤寒沙门氏菌B株的新噬菌体,鸡尾酒的目标范围被转移到包括副伤寒沙门氏菌B株。改良后的鸡尾酒SalmoLyse™能够显著(p < 0.05)减少乙型副伤寒菌株表面污染> 99% (2.1-4.1 log)。数据表明,两种噬菌体鸡尾酒都能有效地显著降低坚硬表面上沙门氏菌的水平,前提是污染菌株在现场试验中敏感(即,在随后的表面去污研究中,现场试验的敏感性表明了有效性)。这些数据还支持噬菌体制剂可以定制以满足所需抗菌应用的概念。
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引用次数: 1
Life in Science: Armin Dale Kaiser. 《科学生活》:阿明·戴尔·凯泽。
Pub Date : 2013-07-01 Epub Date: 2013-06-28 DOI: 10.4161/bact.25589
Armin Dale Kaiser
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引用次数: 0
Innate and acquired bacteriophage-mediated immunity. 先天和获得性噬菌体介导免疫。
Pub Date : 2013-07-01 Epub Date: 2013-07-29 DOI: 10.4161/bact.25857
Jeremy J Barr, Merry Youle, Forest Rohwer

We recently described a novel, non-host-derived, phage-mediated immunity active at mucosal surfaces, the main site of pathogen entry in metazoans. In that work, we showed that phage T4 adheres to mucus glycoproteins via immunoglobulin-like domains displayed on its capsid. This adherence positions the phage in mucus surfaces where they are more likely to encounter and kill bacteria, thereby benefiting both the phage and its metazoan host. We presented this phage-metazoan symbiosis based on an exclusively lytic model of phage infection. Here we extend our bacteriophage adherence to mucus (BAM) model to consider the undoubtedly more complex dynamics in vivo. We hypothesize how mucus-adherent phages, both lytic and temperate, might impact the commensal microbiota as well as protect the metazoan epithelium from bacterial invasion. We suggest that BAM may provide both an innate and an acquired antimicrobial immunity.

我们最近描述了一种新的、非宿主来源的、在粘膜表面活跃的噬菌体介导的免疫,这是后生动物病原体进入的主要部位。在这项工作中,我们发现噬菌体T4通过其衣壳上显示的免疫球蛋白样结构域粘附在粘液糖蛋白上。这种粘附使噬菌体处于粘液表面,在那里它们更有可能遇到并杀死细菌,从而使噬菌体及其后生动物宿主都受益。我们提出了这种噬菌体-后生动物共生基于噬菌体感染的完全溶解模型。在这里,我们扩展了我们的噬菌体黏液粘附(BAM)模型,以考虑无疑更复杂的体内动力学。我们假设黏液粘附噬菌体(裂解性和温带)如何影响共生微生物群以及保护后生动物上皮免受细菌侵袭。我们认为BAM可能同时提供先天和获得性抗微生物免疫。
{"title":"Innate and acquired bacteriophage-mediated immunity.","authors":"Jeremy J Barr,&nbsp;Merry Youle,&nbsp;Forest Rohwer","doi":"10.4161/bact.25857","DOIUrl":"https://doi.org/10.4161/bact.25857","url":null,"abstract":"<p><p>We recently described a novel, non-host-derived, phage-mediated immunity active at mucosal surfaces, the main site of pathogen entry in metazoans. In that work, we showed that phage T4 adheres to mucus glycoproteins via immunoglobulin-like domains displayed on its capsid. This adherence positions the phage in mucus surfaces where they are more likely to encounter and kill bacteria, thereby benefiting both the phage and its metazoan host. We presented this phage-metazoan symbiosis based on an exclusively lytic model of phage infection. Here we extend our bacteriophage adherence to mucus (BAM) model to consider the undoubtedly more complex dynamics in vivo. We hypothesize how mucus-adherent phages, both lytic and temperate, might impact the commensal microbiota as well as protect the metazoan epithelium from bacterial invasion. We suggest that BAM may provide both an innate and an acquired antimicrobial immunity.</p>","PeriodicalId":8686,"journal":{"name":"Bacteriophage","volume":"3 3","pages":"e25857"},"PeriodicalIF":0.0,"publicationDate":"2013-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/bact.25857","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31862802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 66
Properties and mutation studies of a bacteriophage-derived chimeric recombinant staphylolytic protein P128: Comparison to recombinant lysostaphin. 噬菌体衍生的嵌合重组葡萄球菌溶解蛋白P128的性质和突变研究:与重组溶葡萄球菌蛋白的比较。
Pub Date : 2013-07-01 Epub Date: 2013-10-02 DOI: 10.4161/bact.26564
Sanjeev Rajagopalan Saravanan, Vivek Daniel Paul, Shilpa George, Sudarson Sundarrajan, Nirmal Kumar, Madhavi Hebbur, Naveen Kumar, Ananda Veena, Uma Maheshwari, Chemira Biddappa Appaiah, Muralidharan Chidambaran, Anuradha Gopal Bhat, Sukumar Hariharan, Sriram Padmanabhan

P128 is a chimeric anti-staphylococcal protein having a catalytic domain from a Staphylococcus bacteriophage K tail associated structural protein and a cell wall targeting domain from the Staphylococcus bacteriocin-lysostaphin. In this study, we disclose additional properties of P128 and compared the same with lysostaphin. While lysostaphin was found to get inactivated by heat and was inactive on its parent strain S. simulans biovar staphylolyticus, P128 was thermostable and was lytic towards S. simulans biovar staphylolyticus demonstrating a difference in their mechanism of action. Selected mutation studies of the catalytic domain of P128 showed that arginine and cysteine, at 40th and 76th positions respectively, are critical for the staphylolytic activity of P128, although these amino acids are not conserved residues. In comparison to native P128, only the R40S mutant (P301) was catalytically active on zymogram gel and had a similar secondary structure, as assessed by circular dichroism analysis and in silico modeling with similar cell binding properties. Mutation of the arginine residue at 40th position of the P128 molecule caused dramatic reduction in the Vmax (∆OD600 [mg/min]) value (nearly 270 fold) and the recombinant lysostaphin also showed lesser Vmax value (nearly 1.5 fold) in comparison to the unmodified P128 protein. The kinetic parameters such as apparent Km (KmAPP) and apparent Kcat (KcatAPP) of the native P128 protein also showed significant differences in comparison to the values observed for P301 and lysostaphin.

P128是一种嵌合抗葡萄球菌蛋白,其催化结构域来自葡萄球菌噬菌体K尾相关结构蛋白,细胞壁靶向结构域来自葡萄球菌溶葡萄素。在这项研究中,我们揭示了P128的其他特性,并将其与溶葡萄球菌蛋白进行了比较。溶葡萄球菌素在高温下失活,对其亲本菌株拟南葡萄球菌无活性,而P128具有耐热性,对拟南葡萄球菌有裂解作用,两者作用机制不同。P128催化结构域的选择性突变研究表明,精氨酸和半胱氨酸分别位于第40位和第76位,对P128的葡萄球菌降解活性至关重要,尽管这些氨基酸不是保守残基。与天然P128相比,只有R40S突变体(P301)在酶谱凝胶上具有催化活性,并且具有相似的二级结构,通过圆二色性分析和硅模型评估具有相似的细胞结合特性。P128分子第40位精氨酸残基的突变使Vmax(∆OD600 [mg/min])值显著降低(约270倍),重组溶葡萄球菌蛋白的Vmax值也比未修饰的P128蛋白低(约1.5倍)。P128蛋白的表观Km (KmAPP)和表观KcatAPP (KcatAPP)等动力学参数也与P301和溶葡萄蛋白存在显著差异。
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引用次数: 14
Listeria phages: Genomes, evolution, and application. 李斯特菌噬菌体:基因组、进化和应用。
Pub Date : 2013-07-01 Epub Date: 2013-10-24 DOI: 10.4161/bact.26861
Jochen Klumpp, Martin J Loessner

Listeria is an important foodborne pathogen and the causative agent of Listeriosis, a potentially fatal infection. Several hundred Listeria bacteriophages have been described over the past decades, but only few have actually been characterized in some detail, and genome sequences are available for less than twenty of them. We here present an overview of what is currently known about Listeria phage genomics, their role in host evolution and pathogenicity, and their various applications in biotechnology and diagnostics.

李斯特菌是一种重要的食源性病原体,也是李斯特菌病的病原体,李斯特菌病是一种潜在的致命性感染。在过去的几十年中,已经描述了几百种李斯特菌噬菌体,但只有少数噬菌体的特征得到了详细描述,其中只有不到 20 种噬菌体的基因组序列可供使用。我们在此概述目前已知的李斯特菌噬菌体基因组学、它们在宿主进化和致病性中的作用,以及它们在生物技术和诊断学中的各种应用。
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引用次数: 0
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Bacteriophage
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