How does atypical protein kinase C signalling regulate actomyosin-mediated force generation in polarised cells?

非典型蛋白激酶 C 信号传导如何调节极化细胞中肌动球蛋白介导的力产生？

• 批准号: 2608938
• 金额: --
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2608938
• 关键词: How; does; atypical; protein; kinase

Epithelial cells are polarised, and the localisation of the polarity-driving PAR complex overlaps with that of actomyosin significantly. Actomyosin's roles in contractility and cell structure drive morphogenesis during development. The PAR complex effector atypical protein kinase C (aPKC) is linked to actomyosin regulation, yet the full scope of regulatory mechanisms is unclear. aPKC has also been implicated in disease contexts including neurodegeneration and cancer progression. I therefore propose an unbiased screen to identify novel aPKC substrates. Using an analogue-sensitive version of aPKC, substrates will be thiophosphorylated, purified, identified through mass spectrometry, and interactions with aPKC validatedin vitro. Hits will be investigated, with their normal localisation and localisation under acute aPKC inhibition characterised using fluorescence microscopy. Genetic approaches will also be used to elucidate mechanisms driving actomyosin regulation by aPKC. The Drosophila model system will allow analysis of different cell types, enabling the identification of context-dependent aPKC activity.

上皮细胞极化，和本地化的极性驱动PAR复杂的重叠与肌动球蛋白显着。肌动球蛋白在收缩性和细胞结构中的作用驱动发育过程中的形态发生。PAR复合物效应器非典型蛋白激酶C（aPKC）与肌动球蛋白调节有关，但调节机制的全部范围尚不清楚。aPKC还与包括神经退行性疾病和癌症进展在内的疾病背景有关。因此，我提出了一个公正的屏幕，以确定新的aPKC底物。使用aPKC的类似物敏感版本，底物将被硫代磷酸化，纯化，通过质谱鉴定，并在体外验证与aPKC的相互作用。将研究命中，使用荧光显微镜表征其正常定位和急性aPKC抑制下的定位。遗传学方法也将用于阐明驱动肌动球蛋白调节aPKC的机制。果蝇模型系统将允许分析不同的细胞类型，从而能够鉴定上下文依赖性aPKC活性。