Understanding an ancient universal membrane effector system

了解古老的通用膜效应器系统

• 批准号: BB/X003035/1
• 负责人: Gavin Thomas
• 金额: $ 564.72万
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX003035%2F1
• 关键词: Understanding; ancient; universal; membrane; effector

All living cells are surrounded by a thin membrane, which keeps the inside of the cell separate from the outside environment. This essential cellular boundary layer contains proteins that allow the cell to take up food and expel waste products. The membrane is also energised, which means that cells actively generate and maintain ion gradients and voltage across the membrane. This so-called electrochemical gradient is one of the key cellular methods to store energy, and which can be utilised to drive uptake of nutrients and expel waste products. Furthermore, electrochemical gradient is critical for the synthesis of ATP, which is a key molecule used to energise intracellular processes. Hence, the integrity of the membrane is central to the proper function of living cells, and many known toxins including some antibiotics act by disrupting the membrane. During the early evolution of cells, a protein named IM30 evolved that has a role in protecting the membrane in the presence of damaging agents. These proteins, discovered just over 30 year ago, have been found to adopt ring-like structures that can stack upon each other to form tubes. Further, these proteins are known to accumulate in cells to very high levels when their membrane is damaged, and directly bind to cell membranes. However, how they protect the membrane, allowing the electrochemical potential to be maintained, is unknown. In this project we have assembled a diverse team of researchers at different stages in their career and with different sets of expertise. We all share the interest in trying to figure out how the IM30 proteins work to protect cellular membranes. They could potentially do this by forming a coat covering the inside of the membrane, by forming 'ribs' that wrap around the cell and hold the membrane together, or by allowing the membranes to form small fragments that carry away the toxin that is damaging the cell. Alternatively, they might specifically project the proteins that generate the electrochemical potential or use it make ATP. Currently, we simply do not know. Whatever the mechanism is, however, it will be a completely new one and tell us important fundamental information about how biological membranes are organised and function. To figure out how these proteins function, we will use a large number of different microbial species, as these represent relatively simple cells that we can study effectively, and where we know IM30s are important. We will systematically characterise how much of the protein different cells contain, where in the cells the proteins a localised, whether they assemble into rings and rods inside the cell, and how their function is regulated by other factors. We will develop new techniques to measure the electrochemical gradient in real time in living cells, which will provide us with essential tools to study the mechanisms through which IM30 proteins protect cell membranes. Our team comprises of microbiologists, biophysicist, biochemists, geneticists and cell biologists at 5 different Universities in the UK, who will come together to bring about a step change in understanding of IM30 protein function, and more generally how cells protect themselves from environmental insult.

所有活细胞都被一层薄膜包围，使细胞内部与外部环境分开。这个重要的细胞边界层含有蛋白质，使细胞能够吸收食物并排出废物。膜也被通电，这意味着细胞主​​动产生并维持膜上的离子梯度和电压。这种所谓的电化学梯度是细胞储存能量的关键方法之一，可用于促进营养物质的吸收和排出废物。此外，电化学梯度对于 ATP 的合成至关重要，ATP 是用于为细胞内过程提供能量的关键分子。因此，膜的完整性对于活细胞的正常功能至关重要，许多已知的毒素（包括一些抗生素）通过破坏膜来发挥作用。在细胞的早期进化过程中，一种名为 IM30 的蛋白质进化出来，它在存在损伤剂的情况下具有保护细胞膜的作用。这些蛋白质是 30 多年前发现的，被发现采用环状结构，可以相互堆叠形成管状结构。此外，已知当细胞膜受损时，这些蛋白质会在细胞中积累到非常高的水平，并直接与细胞膜结合。然而，它们如何保护膜，从而保持电化学电势尚不清楚。在这个项目中，我们组建了一支多元化的研究人员团队，他们处于职业生涯的不同阶段，具有不同的专业知识。我们都希望弄清楚 IM30 蛋白如何发挥保护细胞膜的作用。他们可能通过形成覆盖细胞膜内部的涂层、形成包裹细胞并将细胞膜固定在一起的“肋”、或者通过让细胞膜形成小碎片来带走损害细胞的毒素来实现这一点。细胞。或者，他们可能会专门投射产生电化学势的蛋白质或使用它来制造 ATP。目前，我们根本不知道。然而，无论其机制是什么，它都将是一种全新的机制，并告诉我们有关生物膜如何组织和发挥作用的重要基本信息。为了弄清楚这些蛋白质的功能，我们将使用大量不同的微生物物种，因为它们代表相对简单的细胞，我们可以有效地研究它们，并且我们知道 IM30 的重要性。我们将系统地描述不同细胞含有多少蛋白质、蛋白质在细胞中的定位、它们是否在细胞内组装成环和杆、以及它们的功能如何受到其他因素的调节。我们将开发新技术来实时测量活细胞中的电化学梯度，这将为我们提供研究 IM30 蛋白保护细胞膜机制的必要工具。我们的团队由来自英国 5 所不同大学的微生物学家、生物物理学家、生物化学家、遗传学家和细胞生物学家组成，他们将齐聚一堂，推动对 IM30 蛋白功能以及更广泛的细胞如何保护自身免受环境侵害的理解发生重大变化。

项目成果

Interrogation of RNA-protein interaction dynamics in bacterial growth

细菌生长中 RNA-蛋白质相互作用动力学的探究

• DOI: 10.1101/2023.07.03.547468
• 发表时间: 2023
• 作者: Monti M

The structural basis of hyperpromiscuity in a core combinatorial network of type II toxin-antitoxin and related phage defense systems.

• DOI: 10.1073/pnas.2305393120
• 发表时间: 2023-08-15
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Ernits, Karin;Saha, Chayan Kumar;Brodiazhenko, Tetiana;Chouhan, Bhanu;Shenoy, Aditi;Buttress, Jessica A.;Duque-Pedraza, Julian J.;Bojar, Veda;Nakamoto, Jose A.;Kurata, Tatsuaki;Egorov, Artyom A.;Shyrokova, Lena;Johansson, Marcus J. O.;Mets, Toomas;Rustamova, Aytan;Dzigurski, Jelisaveta;Tenson, Tanel;Garcia-Pino, Abel;Strahl, Henrik;Elofsson, Arne;Hauryliuk, Vasili;Atkinson, Gemma C.
• 通讯作者: Atkinson, Gemma C.

Flipping the switch: dynamic modulation of membrane transporter activity in bacteria.

• DOI: 10.1099/mic.0.001412
• 发表时间: 2023-11
• 期刊: MICROBIOLOGY-SGM
• 影响因子: 2.8
• 作者: Elston, Rory;Mulligan, Christopher;Thomas, Gavin H.
• 通讯作者: Thomas, Gavin H.