Localised protein synthesis in fibroblasts during cell spreading and migration in 3D culture

3D 培养中细胞扩散和迁移过程中成纤维细胞的局部蛋白质合成

• 批准号: BB/H018956/1
• 负责人: Simon Morley
• 金额: $ 41.91万
• 依托单位: University of Sussex
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH018956%2F1
• 关键词: Localised; protein; synthesis; fibroblasts; during

The ability of cells to move about is an important process in development, generation of blood vessels and repair of damaged tissues after injury, requiring new proteins to be made. For cells to be able to do this, critical information stored in the gene sequences of the genetic material (DNA) has to be decoded by the cell to produce a wide variety of essential proteins of the right type, in the right amount and at the right time. The general transfer of information from DNA to protein is carried out by the messenger RNA (mRNA), which is a copy of the DNA sequence. When required, this mRNA has to be decoded into protein in different parts of the cell by a complex, highly regulated machine termed a ribosome, in a process known as translation. Localised protein synthesis allows the cell to make the protein exactly where and when it needs it in the cell without having to waste time and energy moving the protein around to the correct location. To work efficiently, accurately, and to allow the ribosome to function in the best interests of the cell, this machinery requires helper proteins (translation initiation factors; eIF) that interact with each other, and also make sure that the mRNA and the ribosome come together into a highly regulated, large initiation complex to make the proteins required. So how does the cell make this happen in the right place at the right time? The interaction of the initiation factors themselves is a major site for regulation in mammalian cells. One protein, 4E-binding protein 1 (4E-BP1) prevents the interaction of eIF4E with the scaffold protein, eIF4G, and stops the recruitment of mRNA to the ribosome and halts protein synthesis. When protein synthesis is needed, the cell signals to 4E-BP1 to release the eIF4E/mRNA from the 4E-BP1/eIF4E/mRNA complex to let it work. The cell does this by marking the 4E-BP1, eIF4E, eIF4G and ribosomes with phosphate groups in a process known as phosphorylation. This modification promotes 4E-BP1 release from eIF4E/mRNA which can subsequently bind to eIF4G and form the multi-protein initiation complex required to make the correct types and amounts of protein needed. However, we still do not know how the cell controls localised protein synthesis in cells which are in the process of migrating. From 'looking' inside the cell with specialised microscopy techniques, we know that the initiation factors are discretely localised to specific regions in the cell; they are not just floating about. In the work described here we want to investigate where and how fibroblasts cells localise their translational machinery when they are prompted to migrate. We then want to understand which signals are required to bring about this localisation and show whether these regions reflect active areas where proteins are being made as the cell moves about in culture. These studies will substantially increase our general understanding of the significance of the control of protein synthesis in the regulation of cell growth and migration, opening up new potential avenues for controlling cancer cells which have acquired the ability to move about the body.

细胞移动的能力是发育，血管产生和受伤后受损组织的修复的重要过程，需要制造新的蛋白质。为了使细胞能够做到这一点，必须通过细胞解码存储在遗传物质（DNA）基因序列中的关键信息，以在适当的时间和适当的时间产生正确类型的各种必需蛋白质。信息从DNA到蛋白质的一般传递是由Messenger RNA（mRNA）进行的，该元素是DNA序列的副本。在需要时，必须将该mRNA通过称为核糖体的复合物，高度调节的机器在细胞的不同部分中解码为蛋白质，这是一种称为翻译的过程。局部蛋白质合成使细胞可以使蛋白质在细胞中的何处以及何时在细胞中需要它，而不必浪费时间和能量将蛋白质移动到正确的位置。为了有效，准确，允许核糖体为细胞的最大利益发挥作用，该机械需要相互相互作用的辅助蛋白（翻译起始因子； EIF），并确保mRNA和核糖体与高度调节的大型启动复合物一起制造蛋白质。那么，细胞如何在正确的时间在正确的位置实现这一目标呢？起始因子本身的相互作用本身是哺乳动物细胞调节的主要部位。一种蛋白质，4E结合蛋白1（4E-BP1）可防止EIF4E与脚手架蛋白，EIF4G的相互作用，并阻止mRNA募集到核糖体和停止蛋白质合成中。当需要蛋白质合成时，将细胞信号与4E-BP1信号从4E-BP1/EIF4E/mRNA复合物中释放出EIF4E/mRNA，以使其起作用。该细胞通过在称为磷酸化的过程中标记4E-BP1，EIF4E，EIF4G和核糖体的4E-BP1，EIF4E，EIF4G和核糖体来做到这一点。这种修改促进了从EIF4E/mRNA释放的4E-BP1释放，后来可以与EIF4G结合并形成需要制造正确类型和所需蛋白质的多蛋白起始复合物。但是，我们仍然不知道细胞如何控制迁移过程中细胞中局部蛋白质的合成。从具有专门显微镜技术的细胞内部的“看”，我们知道启动因子被离散地定位于细胞中的特定区域。他们不只是漂浮。在此处描述的工作中，我们想研究成纤维细胞在提示迁移时将其定位的何处以及如何定位。然后，我们想了解需要哪些信号来实现此本地化，并显示这些区域是否反映了在培养物中移动的细胞移动的活跃区域。这些研究将大大提高我们对控制蛋白质合成在细胞生长和迁移调节中的重要性的一般理解，从而为控制癌细胞的新潜在途径开辟了新的潜在途径，这些癌细胞已经获得了围绕身体移动的能力。

项目成果

Murine norovirus 1 (MNV1) replication induces translational control of the host by regulating eIF4E activity during infection.

• DOI: 10.1074/jbc.m114.602649
• 发表时间: 2015-02-20
• 期刊: The Journal of biological chemistry
• 作者: Royall E;Doyle N;Abdul-Wahab A;Emmott E;Morley SJ;Goodfellow I;Roberts LO;Locker N
• 通讯作者: Locker N

Translation initiation factors and active sites of protein synthesis co-localize at the leading edge of migrating fibroblasts

翻译起始因子和蛋白质合成活性位点共定位于迁移成纤维细胞的前缘

• DOI: 10.1042/bj20110435
• 发表时间: 2011
• 期刊: Biochemical Journal
• 影响因子: 4.1
• 作者: Willett M

The helicase, DDX3X, interacts with poly(A)-binding protein 1 (PABP1) and caprin-1 at the leading edge of migrating fibroblasts and is required for efficient cell spreading.

• DOI: 10.1042/bcj20170354
• 发表时间: 2017-08-30
• 期刊: The Biochemical journal
• 作者: Copsey AC;Cooper S;Parker R;Lineham E;Lapworth C;Jallad D;Sweet S;Morley SJ
• 通讯作者: Morley SJ

Sumoylation of eIF4A2 affects stress granule formation.

• DOI: 10.1242/jcs.184614
• 发表时间: 2016-06-15
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Jongjitwimol J;Baldock RA;Morley SJ;Watts FZ
• 通讯作者: Watts FZ