Moving metals: Systematic characterization of metal transporters in Toxoplasma gondii

细胞需要金属来产生能量、生长和分裂。然而，金属不能穿过我们细胞周围的膜，而是通过一种叫做转运蛋白的特殊蛋白质在膜上运输。我们体内的细胞和感染我们的病原体都是如此。我对一种叫做刚地弓形虫的单细胞寄生虫很感兴趣。这种寄生虫可以生活在我们体内任何不同的细胞或组织中，这使得它非常善于适应环境。我们知道弓形虫需要金属，但我们不知道它们是如何将这些金属从我们的细胞运送到寄生虫体内的。在这个项目中，我们将鉴定寄生虫转运蛋白，并研究它们如何在弓形虫中起作用。为了做到这一点，我们将首先研究一些我们所知道的金属转运蛋白，因为它们与在其他生物中研究的金属转运蛋白相似。我们会发现移除这些转运蛋白后会发生什么，以及它们在细胞中的位置。但我们预测有一些类型的转运蛋白与我们所知道的转运蛋白不同。找到这些更有挑战性。我们将收集每个转运蛋白（约200个）被删除的寄生虫。我们将在不同的条件下（例如，不同水平的金属）种植这些寄生虫。通过测量它们的生长量，我们可以找到转运体，当我们删除它们时，它们会改变寄生虫在不同金属条件下生长的能力。这将帮助我们确定在不同的环境中哪些转运蛋白对寄生虫是重要的。为了研究这些转运蛋白的功能，我们还将采用其他方法。我们将看看我们的寄生虫转运体是否能取代那些在酵母中得到充分研究的转运体。如果它们可以，这意味着它们可能具有相同或类似的功能。我们还将把我们的转运器放入青蛙卵中。青蛙卵很特别，因为它们自己没有任何转运体，所以利用这些大细胞，我们可以非常精确地测量寄生虫转运体是如何将金属运入和运出细胞的。在这个项目中，我们将了解弓形虫金属转运体的身份及其在这些寄生虫中的作用。因为金属对寄生虫至关重要，如果我们了解了这一点，我们就能更多地了解寄生虫是如何产生能量的，以及它是如何与细胞或我们的身体相互作用的。这些知识将帮助我们在未来发现治疗弓形虫的新方法，可能还有与之相关的引起疟疾的寄生虫疟原虫。

Cells require metals to create energy and to grow and divide. However, metals cannot pass through the membranes that surround our cells, and instead are transported across membranes by specialised proteins called transporters. This is true of the cells in our bodies, as well as the pathogens which infect us. I am interested in a single-celled parasite called Toxoplasma gondii. This parasite can live in any of the different cells or tissues in our body, making it exceptionally good at adapting to its environment. We know that Toxoplasma requires metals, but we don't know how they are transporting these metals from our cells into the parasite. In this project, we will identify parasite transporter proteins, and investigate how they are functioning within Toxoplasma. To do this, we will first investigate some metal transporters that we know about because they are similar to ones studied in other organisms. We will find out what happens when we remove these transporters, and where they are localised on the cell. But we predict there are types of transporter proteins that are not similar to those we know about. To find these is more challenging. We will make a collection of parasites with every transporter (about 200) deleted. We will grow this collection of parasites in different conditions (e.g. with different levels of metals). By measuring how much they grow, we can find transporters that, when we delete them, change the parasite's ability to grow under different metal conditions. This will help us identify which transporters are important to the parasite in different environments.To investigate how these transporter proteins function, we will also take other approaches. We will see if our parasite transporters can replace ones that are well-studied in yeast. If they can it means that they are likely to have the same or a similar function. We will also put our transporters into frog eggs. Frog eggs are special because they do not have any transporters of their own, so using these large cells we can measure very precisely how our parasite transporters are working to move metals into and out of the cell.In this project, we will learn the identity of Toxoplasma metal transporters and how they function in these parasites. Because metals are essential to the parasite, if we understand this we will understand more about how the parasite produces energy and how it interacts with the cells or our body. This knowledge will help us in the future discover new way to treat Toxoplasma, and possibly its related parasite Plasmodium, which causes malaria.