Department of the Geophysical Sciences, University of Chicago, Chicago, IL 60637
Department of the Geophysical Sciences, University of Chicago, Chicago, IL 60637.
Proc Natl Acad Sci U S A. 2019 Jul 30;116(31):15590-15595. doi: 10.1073/pnas.1901856116. Epub 2019 Jul 15.
The building blocks of a virus derived from de novo biosynthesis during infection and/or catabolism of preexisting host cell biomass, and the relative contribution of these 2 sources has important consequences for understanding viral biogeochemistry. We determined the uptake of extracellular nitrogen (N) and its biosynthetic incorporation into both virus and host proteins using an isotope-labeling proteomics approach in a model marine cyanobacterium WH8102 infected by a lytic cyanophage S-SM1. By supplying dissolved N as N postinfection, we found that proteins in progeny phage particles were composed of up to 41% extracellularly derived N, while proteins of the infected host cell showed almost no isotope incorporation, demonstrating that de novo amino acid synthesis continues during infection and contributes specifically and substantially to phage replication. The source of N for phage protein synthesis shifted over the course of infection from mostly host derived in the early stages to more medium derived later on. We show that the photosystem II reaction center proteins D1 and D2, which are auxiliary metabolic genes (AMGs) in the S-SM1 genome, are made de novo during infection in an apparently light-dependent manner. We also identified a small set of host proteins that continue to be produced during infection; the majority are homologs of AMGs in S-SM1 or other viruses, suggesting selective continuation of host protein production during infection. The continued acquisition of nutrients by the infected cell and their utilization for phage replication are significant for both evolution and biogeochemical impact of viruses.
病毒的构建块是在感染过程中从头合成的,或者是从前宿主细胞生物质的分解代谢中产生的,这两种来源的相对贡献对于理解病毒生物地球化学具有重要意义。我们使用同位素标记蛋白质组学方法,在一种模式海洋蓝藻 WH8102 中感染裂解性蓝藻噬菌体 S-SM1 后,确定了细胞外氮(N)的摄取及其在病毒和宿主蛋白质中的生物合成掺入。通过在感染后提供溶解 N 作为 N 的来源,我们发现,在子代噬菌体颗粒中的蛋白质中,最多有 41%的 N 是由细胞外衍生而来的,而受感染的宿主细胞中的蛋白质几乎没有同位素掺入,这表明从头氨基酸合成在感染过程中持续进行,并专门且大量地有助于噬菌体复制。在感染过程中,噬菌体蛋白合成的 N 来源从早期的主要来源于宿主转变为后期更多的来源于培养基。我们表明,在感染过程中,光合作用系统 II 反应中心蛋白 D1 和 D2 以明显依赖光的方式从头合成,这些蛋白是 S-SM1 基因组中的辅助代谢基因(AMGs)。我们还鉴定了一小部分在感染过程中继续产生的宿主蛋白;大多数是 S-SM1 或其他病毒中的 AMGs 的同源物,这表明在感染过程中宿主蛋白的产生具有选择性。受感染细胞持续获取营养物质并将其用于噬菌体复制,这对病毒的进化和生物地球化学影响都具有重要意义。