How do astrotactin-1 and astrotactin-2 act in the determination of mammalian cell polarity?

astrotactin-1 和 astrotactin-2 在确定哺乳动物细胞极性中如何发挥作用？

• 批准号: MR/N000331/1
• 负责人: Robert Gilbert
• 金额: $ 40.2万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FN000331%2F1
• 关键词: How; do; astrotactin; act; determination

The growth, development and repair of the brain requires the migration of neurons so they can shape its sub-structures, make new connections, and repair old ones. Similarly, the growth of other tissues and organs requires the growth of cells in a polarised fashion, to give the distinctive shapes characteristic of body plans and individual organs within them. It has recently been found that a pair of proteins called astrotactins (ASTNs) play critical roles in both these processes.In the brain, ASTN1 is found on the surface of neurons where it makes connections to the guide tracks ("glial fibres") along which they move. In order for the neurons to move, however, they need to break old connections and then form new ones, and in order to do that ASTN1-mediated contact sites need to be recycled towards the leading edge of the migrating cell. The key molecule enabling that to happen is ASTN2, which is a very similar protein to ASTN1 but which is found in small membrane-defined compartments within the cell rather than sticking out of the cell membrane at its surface, like ASTN1 does. In other tissues, our collaborators at Johns Hopkins University School of Medicine in the United States recently found that ASTN2 helps control something known as planar cell polarity, which underpins the formation of organs and tissues both in the nervous system and elsewhere in the body.Clinical and genetic data indicate a key involvement of mutant forms of ASTN1 and ASTN2 in a host of conditions including attention deficit-hyperactivity disorder (ADHD), autism spectrum disorders, schizophrenia and forms of neurodevelopmental delay such as difficulty in learning language. Another recent study showing that different forms of ASTN2 could bring forward by ~5 years the age of onset of Alzheimer's, while effects of ASTNs have also been noted on the immune system and general tissue morphology outside the nervous system.We are going to use methods which give us a 3D atomic-resolution picture of the structure of ASTN1 and ASTN2 to work out how they carry out their activities controlling normal human development, repair when tissues are damaged, and the suceptibility to conditions like Alzheimer's. Preliminary data from our lab and others suggests that what is critical is a change in the behaviour of ASTN1 and ASTN2 in more acidic conditions - different compartments within the cell have differing acidities and this regulates events within them. We have an excellent starting point because we have crystallised ASTN1 in two different forms, at neutral pH and at acidic pH. We have also made preliminary studies of how they interact but will be able to provide much greater detail from the work we are planning now.Clearly, we also need to look at living cells and see how ASTN1 and ASTN2 move about within them, affecting each other's activity and location. We will do this as well, which will give a functional context to the work we are doing on the atomic structures of the proteins. The approach we will use is based on fluorescently labelling the proteins and this will enable us both to see the location of the ASTN proteins and to measure the interactions they undergo.Our work will benefit greatly from ongoing collaboration both with the group at Johns Hopkins and with colleagues local to Oxford. One particularly exciting area to explore further will be the way in which ASTN1 and ASTN2 interact with proteins already known to play leading roles in determining planar call polarity, such as the cell surface receptor proteins Frizzled-3 and Frizzled-6 and Celsr1.

大脑的生长，发育和修复需要神经元的迁移，以便它们能够塑造其子结构，建立新的连接并修复旧的连接。类似地，其他组织和器官的生长需要细胞以极化的方式生长，以给出身体计划和其中单个器官的独特形状。最近发现一对被称为astrotactins（ASTN）的蛋白质在这两个过程中起着关键作用。在大脑中，ASTN 1被发现存在于神经元的表面，在那里它与它们移动的导向轨道（“胶质纤维”）沿着连接。然而，为了使神经元移动，它们需要打破旧的连接，然后形成新的连接，为了做到这一点，ASTN 1介导的接触位点需要向迁移细胞的前沿回收。实现这一目标的关键分子是ASTN 2，它是一种与ASTN 1非常相似的蛋白质，但它存在于细胞内的小的膜限定区室中，而不是像ASTN 1那样从细胞膜表面伸出。在其他组织中，我们在美国约翰霍普金斯大学医学院的合作者最近发现，ASTN 2有助于控制被称为平面细胞极性的东西，临床和遗传数据表明，ASTN 1和ASTN 2的突变形式在包括注意力缺陷在内的许多疾病中起着关键作用。多动症（ADHD）、自闭症谱系障碍、精神分裂症和神经发育迟缓形式如学习语言困难。另一项最近的研究表明，不同形式的ASTN 2可以使阿尔茨海默氏症的发病年龄提前5年，而ASTN的影响也已经注意到对免疫系统和神经系统外的一般组织形态。我们将使用的方法，给我们一个3D原子-ASTN 1和ASTN 2结构的高分辨率图片，以了解它们如何进行控制正常人类发育的活动，当组织受损时进行修复，以及易患老年痴呆症等疾病的风险。来自我们实验室和其他实验室的初步数据表明，关键是ASTN 1和ASTN 2在酸性条件下的行为发生变化-细胞内的不同隔室具有不同的酸度，这调节了其中的事件。我们有一个很好的起点，因为我们已经将ASTN 1在中性pH和酸性pH下以两种不同的形式结晶。我们还对它们如何相互作用进行了初步研究，但我们现在计划的工作将能够提供更详细的信息。显然，我们还需要观察活细胞，看看ASTN 1和ASTN 2如何在其中移动，影响彼此的活动和位置。我们也会这样做，这将为我们在蛋白质原子结构上所做的工作提供一个功能背景。我们将使用的方法是基于荧光标记的蛋白质，这将使我们既可以看到的ASTN蛋白质的位置，并测量它们所经历的相互作用。我们的工作将大大受益于正在进行的合作，无论是与该集团在约翰霍普金斯和当地的同事到牛津大学。一个特别令人兴奋的领域将进一步探索ASTN 1和ASTN 2与已知在确定平面调用极性中发挥主导作用的蛋白质相互作用的方式，例如细胞表面受体蛋白Frizzled-3和Frizzled-6和Celsr 1。

项目成果

Repurposing a pore: highly conserved perforin-like proteins with alternative mechanisms.

• DOI: 10.1098/rstb.2016.0212
• 发表时间: 2017-08-05
• 期刊: Philosophical transactions of the Royal Society of London. Series B, Biological sciences
• 作者: Ni T;Gilbert RJC
• 通讯作者: Gilbert RJC

Cryo-EM structures of perforin-2 in isolation and assembled on a membrane suggest a mechanism for pore formation.

• DOI: 10.15252/embj.2022111857
• 发表时间: 2022-12-01
• 期刊: The EMBO journal

Structural, Functional and Computational Studies of Membrane Recognition by Plasmodium Perforin-Like Proteins 1 and 2.

疟原虫穿孔素样蛋白 1 和 2 膜识别的结构、功能和计算研究。

• DOI: 10.1016/j.jmb.2022.167642
• 发表时间: 2022
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Williams SI

Structures of monomeric and oligomeric forms of the Toxoplasma gondii perforin-like protein 1.

• DOI: 10.1126/sciadv.aaq0762
• 发表时间: 2018-03
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Ni T;Williams SI;Rezelj S;Anderluh G;Harlos K;Stansfeld PJ;Gilbert RJC
• 通讯作者: Gilbert RJC

Structure of astrotactin-2: a conserved vertebrate-specific and perforin-like membrane protein involved in neuronal development.

• DOI: 10.1098/rsob.160053
• 发表时间: 2016-05
• 期刊: Open biology
• 影响因子: 5.8
• 作者: Ni T;Harlos K;Gilbert R
• 通讯作者: Gilbert R