Functions and interactions of the Rad6 pathway in vertebrates

Rad6上位性组通过诱变性跨损伤聚合酶或基于重组的无错误修复模式控制酵母复制期间DNA损伤的旁路。Rad6途径的成员在高等真核生物中是保守的，但迄今为止其功能的确切保守程度尚不清楚。靶向诱变和重组在免疫系统中的抗体多样化中起主要作用，但在这些过程中如何调节修复途径决定和易错聚合酶的募集尚不清楚。我们已经发现，Rad6途径参与了鸡B细胞系DT 40中免疫球蛋白基因的体细胞超突变，这表明该途径还控制了脊椎动物抗体多样化过程中诱变和重组之间的决定。在本项目中，我们希望更详细地研究Rad6途径在脊椎动物细胞中的一般以及靶向同源重组和诱变过程中的功能。此外，我们希望采用亲和纯化来研究Rad6途径成员之间的相互作用以及与其他DNA修复和细胞周期控制途径的相互作用。

The Rad6 epistasis group controls the bypass of DNA lesions during replication in yeast by either mutagenic translesion polymerases or by recombination based error free repair modes. Members of the Rad6 pathway are conserved in higher eukaryotes, but the exact degree of conservation of their function is unclear to date. Targeted mutagenesis and recombination play a major role in antibody diversification in the immune system, but it is unknown how repair pathway decision and recruitment of error-prone polymerases is regulated during these processes. We have found that the Rad6 pathway is involved in somatic hypermutation of immunoglobulin genes in the chicken B cell line DT40, suggesting that this pathway also controls the decision between mutagenesis and recombination during antibody diversification in vertebrates. In the present project, we wish to investigate in more detail the functions of the Rad6 pathway during general as well as targeted homologous recombination and mutagenesis in vertebrate cells. Also, we want to employ affinity purification to study the interactions of Rad6 pathway members with each other and with other pathways of DNA repair and cell cycle control.