Phosphorylation-mediated regulation of the NF-kB subunit c-Rel

核因子κ B （NF-kB）是免疫、应激和炎症反应的中枢调节因子，在细胞分化和存活中也起着关键作用。NF-kB的异常调节也与许多病理状况有关，包括炎症性疾病和癌症。质谱和细胞生物学技术将用于鉴定和表征B细胞中c-Rel NF-kB亚基的翻译后修饰（PTMs）的功能。生物质谱法将用于PTM位点鉴定、蛋白质定量和自上而下的蛋白质组学。基于生物化学/细胞生物学的方法将用于阐明所发现的c-Rel修饰位点的生物学作用/转录介导。这包括细胞培养、基于细胞核转染的b细胞淋巴瘤细胞系、Western blotting、c-Rel靶基因表达的q-PCR分析和细胞活力分析。随着项目的进展和对c-Rel及其相互作用的更多信息的获取，将使用c-Rel突变表达质粒进行功能实验，以更深入地了解c-Rel的作用。

Nuclear Factor kappa B (NF-kB) is a central regulator of the immune, stress and inflammatory responses, and additionally plays key roles in cell differentiation and survival. Aberrant regulation of NF-kB is also associated with many pathological conditions, including inflammatory diseases and cancer.Mass spectrometric and cell biological techniques will be used to identify and characterise the functions of post-translational modifications (PTMs) of the c-Rel NF-kB subunit in B cells. Biological mass spectrometry will be used in PTM site identification, protein quantification and top-down proteomics. Biochemical/cell biology based approaches will be used to elucidate the biological role/transcription mediation of the c-Rel modification sites that are found. This includes cell culture, nucleofection-based transfection of B-cell lymphoma cell lines, Western blotting, q-PCR analysis of c-Rel target gene expression and analysis of cell viability. As the project progresses and more information is acquired about c-Rel and its interactions, functional experiments using c-Rel mutant expression plasmids will be used in order to gain more insight into the roles of c-Rel.