Department of Microbiology, Immunology and Molecular Genetics, UT Health, San Antonio, TX 78229, USA.
Department of Biochemistry and Structural Biology, UT Health, San Antonio, TX 78229, USA.
Int J Mol Sci. 2023 May 18;24(10):8941. doi: 10.3390/ijms24108941.
Diversity of phage propagation, physical properties, and assembly promotes the use of phages in ecological studies and biomedicine. However, observed phage diversity is incomplete. siphophage, 0105phi-7-2, first described here, significantly expands known phage diversity, as seen via in-plaque propagation, electron microscopy, whole genome sequencing/annotation, protein mass spectrometry, and native gel electrophoresis (AGE). Average plaque diameter vs. plaque-supporting agarose gel concentration plots reveal unusually steep conversion to large plaques as agarose concentration decreases below 0.2%. These large plaques sometimes have small satellites and are made larger by orthovanadate, an ATPase inhibitor. Phage head-host-cell binding is observed by electron microscopy. We hypothesize that this binding causes plaque size-increase via biofilm evolved, ATP stimulated ride-hitching on motile host cells by temporarily inactive phages. Phage 0105phi7-2 does not propagate in liquid culture. Genomic sequencing/annotation reveals history as temperate phage and distant similarity, in a virion-assembly gene cluster, to prototypical siphophage SPP1 of . Phage 0105phi7-2 is distinct in (1) absence of head-assembly scaffolding via either separate protein or classically sized, head protein-embedded peptide, (2) producing partially condensed, head-expelled DNA, and (3) having a surface relatively poor in AGE-detected net negative charges, which is possibly correlated with observed low murine blood persistence.
噬菌体的繁殖方式、物理特性和组装方式多种多样,这促进了噬菌体在生态研究和生物医学中的应用。然而,观察到的噬菌体多样性并不完整。本文首次描述的 siphophage 0105phi-7-2 极大地扩展了已知噬菌体的多样性,这可以通过在斑块中繁殖、电子显微镜观察、全基因组测序/注释、蛋白质质谱分析和天然凝胶电泳(AGE)来观察到。平均斑块直径与支持斑块的琼脂糖凝胶浓度的关系图显示,当琼脂糖浓度降低到 0.2%以下时,会异常陡峭地转化为大斑块。这些大斑块有时会有小卫星,并且通过对磷酸腺苷(ATP)酶的抑制剂偏钒酸钠的作用而变得更大。通过电子显微镜观察到噬菌体头部与宿主细胞的结合。我们假设这种结合通过暂时不活跃的噬菌体在运动宿主细胞上通过生物膜进化的、ATP 刺激的搭便车行为,导致斑块大小的增加。噬菌体 0105phi7-2 不能在液体培养中繁殖。基因组测序/注释显示,它具有温和噬菌体的历史,并且在病毒组装基因簇中与典型的 siphophage SPP1 of 具有遥远的相似性。噬菌体 0105phi7-2 在以下方面具有独特性:(1)没有通过单独的蛋白质或经典大小的头部蛋白嵌入肽来提供头部组装支架;(2)产生部分浓缩的、头部排出的 DNA;(3)表面在 AGE 检测到的净负电荷方面相对较差,这可能与观察到的低鼠血持久性相关。