Pope Welkin H, Weigele Peter R, Chang Juan, Pedulla Marisa L, Ford Michael E, Houtz Jennifer M, Jiang Wen, Chiu Wah, Hatfull Graham F, Hendrix Roger W, King Jonathan
Department of Biology, Massachusetts Institute of Technology, Cambridge MA 02139, USA.
J Mol Biol. 2007 May 11;368(4):966-81. doi: 10.1016/j.jmb.2007.02.046. Epub 2007 Feb 22.
Marine Synechococcus spp and marine Prochlorococcus spp are numerically dominant photoautotrophs in the open oceans and contributors to the global carbon cycle. Syn5 is a short-tailed cyanophage isolated from the Sargasso Sea on Synechococcus strain WH8109. Syn5 has been grown in WH8109 to high titer in the laboratory and purified and concentrated retaining infectivity. Genome sequencing and annotation of Syn5 revealed that the linear genome is 46,214 bp with a 237 bp terminal direct repeat. Sixty-one open reading frames (ORFs) were identified. Based on genomic organization and sequence similarity to known protein sequences within GenBank, Syn5 shares features with T7-like phages. The presence of a putative integrase suggests access to a temperate life cycle. Assignment of 11 ORFs to structural proteins found within the phage virion was confirmed by mass-spectrometry and N-terminal sequencing. Eight of these identified structural proteins exhibited amino acid sequence similarity to enteric phage proteins. The remaining three virion proteins did not resemble any known phage sequences in GenBank as of August 2006. Cryo-electron micrographs of purified Syn5 virions revealed that the capsid has a single "horn", a novel fibrous structure protruding from the opposing end of the capsid from the tail of the virion. The tail appendage displayed an apparent 3-fold rather than 6-fold symmetry. An 18 A resolution icosahedral reconstruction of the capsid revealed a T=7 lattice, but with an unusual pattern of surface knobs. This phage/host system should allow detailed investigation of the physiology and biochemistry of phage propagation in marine photosynthetic bacteria.
海洋聚球藻属和海洋原绿球藻属是公海中数量占优势的光合自养生物,也是全球碳循环的贡献者。Syn5是从马尾藻海分离得到的一种短尾蓝藻噬菌体,其宿主为聚球藻菌株WH8109。Syn5已在实验室中于WH8109中培养至高滴度,并进行了纯化和浓缩,且保留了感染性。Syn5的基因组测序和注释显示,其线性基因组为46,214 bp,具有237 bp的末端直接重复序列。共鉴定出61个开放阅读框(ORF)。基于基因组组织以及与GenBank中已知蛋白质序列的序列相似性,Syn5与T7样噬菌体具有共同特征。推测存在整合酶表明其可能具有温和的生命周期。通过质谱分析和N端测序证实了11个ORF与噬菌体病毒粒子中的结构蛋白相关。其中8种鉴定出的结构蛋白与肠道噬菌体蛋白具有氨基酸序列相似性。截至2006年8月,其余三种病毒粒子蛋白与GenBank中任何已知的噬菌体序列均不相似。纯化的Syn5病毒粒子的冷冻电子显微镜照片显示,衣壳有一个单一的“角”,这是一种从病毒粒子尾部相对的衣壳末端突出的新型纤维结构。尾附属物显示出明显的三重而非六重对称性。衣壳的18 Å分辨率二十面体重建显示为T = 7晶格,但表面瘤的模式不同寻常。这种噬菌体/宿主系统将有助于详细研究噬菌体在海洋光合细菌中繁殖的生理学和生物化学。