Exploring transcription of a large DNA virus of importance for global food security

最近的covid-19大流行使病毒成为公众关注的焦点。然而，令人震惊的是，我们对病毒的工作原理知之甚少，包括病毒基因组在转录过程中的控制方式。作为“基因组守门人”的酶被称为RNA聚合酶（RNAP）。rnap是分子机器，可以比作我们日常生活中的其他机器，如汽车。在较小的尺度上，汽车是由由小部件组成的发动机提供动力的，为了使汽车的运动合理化，需要了解它们的形状和功能。虽然这对于了解汽车的机械结构和驾驶过程很重要，但它并不能提供有关交通的信息。在更大的范围内，许多不同类型的车辆共享道路，并在交通规则的指导下以协调的方式在繁忙的城市中行驶。在最大规模上，城市通过一个复杂的网络相互连接，使旅行成为可能。同样，rnap是由许多亚基组成的生命分子机器，这些亚基被整合到高阶转录复合物中。为了了解它们是如何工作的，我们需要确定它们的结构和结构，并在试管中严格控制的条件下描述它们的功能。但在体内，在细胞中，rnap不是单独存在的，而是相互作用，并与不同的机器（包括复制我们基因组的机器）相互作用，这将影响转录的动力学，rnap如何在基因组的背景下分布和功能。这项提议旨在发现病毒RNAP的地图、机制和“交通规则”。对RNAP转录的变革性、详细而全面的理解要求我们在从原子到基因组到细胞水平的一系列生物学尺度上研究这一过程，这需要专门的多学科和多标量研究策略。在我的职业生涯中，我指导了基础研究，利用我的综合多学科研究理念，改变了古生菌转录领域。最近，我的团队首次描述了在病毒-宿主军备竞赛中对古细菌RNAP转录的抑制。我们在Pirbright研究所的合作伙伴Linda Dixon和Chris Netherton是ASFV病毒学领域的全球领导者，我们是开展这项突破性研究的完美团队。

The recent covid-19 pandemic has brought viruses to the forefront of public attention. Yet it is shocking how little we know about how viruses work, including the way by which their genomes are controlled in a process called transcription. The enzymes that serve as the 'gatekeepers of the genome' are called RNA polymerases, RNAP. RNAPs are molecular machines which can be likened to other machines in our daily lives such as cars. At smaller scales, cars are powered by engines that are made of small parts, and to rationalise how cars move requires knowledge about their shape and function. While this is important to understand the mechanics of the car and the process of driving, it does not provide information about traffic. Towards larger scales, many different types of vehicles share the roads and move in a coordinated fashion in a busy city guided by traffic rules. At the largest scale, cities are interconnected by a complex network that enables travel. Likewise, RNAPs are molecular machines of life that consist of many subunits that are incorporated into higher-order transcription complexes. To understand how they work, we need to determine their structure and architecture, and characterise their function under rigorously controlled conditions in vitro, in the test tube. But in vivo, in the cell, RNAPs are not alone but interact with each other and with different machines (including the ones that replicate our genomes) and this will affect the dynamics of transcription, how RNAPs are distributed and function in the context of the genome. This proposal aims to discover the maps, the mechanics, and the 'traffic rules' of a virus RNAP. A transformative, detailed yet comprehensive, understanding of RNAP transcription requires that we study the process over a range of biological scales from the atomic- via the genomic- to the cellular level, which requires a dedicated multidisciplinary and multiscalar research strategy. I have during my career directed fundamental research which has transformed the field of transcription in the Archaea using my brand of integrative multidisciplinary research philosophy. Recently my team characterised, for the first time, the inhibition of archaeal RNAP transcription in the virus-host arms race. Our collaborators Linda Dixon and Chris Netherton at the Pirbright Institute are global leaders in ASFV virology, together we are the perfect team to carry out this ground-breaking study.