Characterisation of the hSNM1B/Apollo protein in the cellular response to DNA damage in the context of the FA/BRCA pathway

FA/BRCA 通路背景下细胞对 DNA 损伤反应中 hSNM1B/Apollo 蛋白的表征

• 批准号: 269195109
• 负责人: Professor Dr. Ilja Demuth
• 金额: --
• 依托单位: Medizinische Klinik für Endokrinologie und Stoffwechselmedizin (Diabetes und Ernährungsmedizin)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/269195109?language=en
• 关键词: Characterisation; hSNM1B; Apollo; protein; cellular

We have identified the hSNM1B/Apollo gene based on its similarity to a yeast (S.cerevisiae) DNA-repair gene, PSO2. The cellular phenotype of cultured cells depleted for hSNM1B/Apollo (siRNA) is reminescent to the cellular phenotype of cells from patients affected from recessive inherited disease Fanconi Anemia (FA). We therefore proposed a hSNM1B/Apollo role within the FA/BRCA pathway in the response to DNA damage. Homozygous mutations in 16 different genes can cause FA, whereas heterozygous mutations in some of these genes are associated with an increased risc to develop cancer (e.g. BRCA2).Recently we were able to establish a direct link between hSNM1B/Apollo and the FA/BRCA pathway: hSNM1B/Apollo interacts physically with one of the known FA proteins, FANCP/SLX4. Here we would like to further narrow down the protein domains in both proteins relevant for this interaction and characterize them in hSNM1B/Apollo null mutant cell lines. Another important aspect of this project will be the analysis of the impact of different SNPs located at the hSNM1B/Apollo locus and known to influence the amount of cellular hSNM1B/Apollo-mRNA on the cellular DNA damage response phenotype. Lymphoblastoid cells of different SNP genotypes will be analysed with respect to mutagen sensitivity and phosphorylation status of relevant proteins (ATM, SMC1) in response to DNA damage.

我们已经确定了hSNM 1B/阿波罗基因的基础上，其相似性酵母（酿酒酵母）DNA修复基因，PSO 2。去除hSNM 1B/Apollo（siRNA）的培养细胞的细胞表型与来自受隐性遗传疾病范可尼贫血（FA）影响的患者的细胞的细胞表型相似。因此，我们提出了hSNM 1B/Apollo在FA/BRCA通路中对DNA损伤的反应中的作用。16个不同基因的纯合突变可导致FA，而其中一些基因的杂合突变与增加的风险相关，从而发展为癌症（例如BRCA 2）。最近，我们能够建立hSNM 1B/Apollo和FA/BRCA通路之间的直接联系：hSNM 1B/Apollo与已知FA蛋白之一FANCP/SLX 4发生物理相互作用。在这里，我们想进一步缩小这两种蛋白质中与这种相互作用相关的蛋白质结构域，并在hSNM 1B/Apollo无效突变细胞系中表征它们。该项目的另一个重要方面将是分析位于hSNM 1B/Apollo位点的不同SNP的影响，已知这些SNP会影响细胞hSNM 1B/Apollo-mRNA对细胞DNA损伤反应表型的量。将分析不同SNP基因型的淋巴母细胞样细胞对诱变剂的敏感性和响应DNA损伤的相关蛋白质（ATM、SMC 1）的磷酸化状态。