Biobizkaia Health Research Institute, Barakaldo, Spain.
Institute for Cancer Research, Oslo University Hospital, The Norwegian Radium Hospital, Oslo, Norway.
Methods Mol Biol. 2024;2743:1-19. doi: 10.1007/978-1-0716-3569-8_1.
Nonsense mutations generating premature termination codons (PTCs) in various genes are frequently associated with somatic cancer and hereditary human diseases since PTCs commonly generate truncated proteins with defective or altered function. Induced translational readthrough during protein biosynthesis facilitates the incorporation of an amino acid at the position of a PTC, allowing the synthesis of a complete protein. This may evade the pathological effect of the PTC mutation and provide new therapeutic opportunities. Several protein tyrosine phosphatases (PTPs) genes are targeted by PTC in human disease, the tumor suppressor PTEN being the more prominent paradigm. Here, using PTEN and laforin as examples, two PTPs from the dual-specificity phosphatase subfamily, we describe methodologies to analyze in silico the distribution and frequency of pathogenic PTC in PTP genes. We also summarize laboratory protocols and technical notes to study the induced translational readthrough reconstitution of the synthesis of PTP targeted by PTC in association with disease in cellular models.
无意义突变在各种基因中产生过早终止密码子 (PTCs)，常与体细胞癌和遗传性人类疾病相关，因为 PTC 通常会产生具有缺陷或改变功能的截短蛋白。在蛋白质生物合成过程中诱导翻译通读有助于在 PTC 的位置掺入一个氨基酸，从而允许合成完整的蛋白质。这可能会规避 PTC 突变的病理影响，并提供新的治疗机会。几种蛋白酪氨酸磷酸酶 (PTP) 基因是人类疾病中 PTC 的靶点，肿瘤抑制因子 PTEN 是更为突出的范例。在这里，我们使用 PTEN 和 laforin 作为例子，两种来自双特异性磷酸酶亚家族的 PTP，描述了分析 PTP 基因中致病性 PTC 分布和频率的计算方法。我们还总结了实验室方案和技术说明，以研究与疾病相关的 PTP 靶向 PTC 在细胞模型中诱导翻译通读重建的情况。
