STRUCTURE OF REV1/PCNA COMPLEX IN PEG

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的目标是确定与DNA聚合酶Rev1的结构域形成的复合体中增殖细胞核抗原的三维结构。Rev1是跨损伤合成DNA聚合酶的一员，允许复制通过DNA损伤，使其成为真核细胞DNA损伤诱导突变的关键因素。最近的研究表明，Rev1在DNA损伤中的高度处理需要它与增殖细胞核抗原的相互作用。研究还表明，这种相互作用在DNA损伤反应过程中是稳定的。与这些研究一致，我们的体外结合分析使用量热法和核磁共振波谱表明，增殖细胞核抗原与位于N端Rev1的BRCT结构域之间存在紧密的相互作用。我们得到了适合于结构测定的PCNA/Rev1-BRCT复合体晶体。用富含硒蛋氨酸的增殖细胞核抗原收集MAD或SAD数据。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our objective is to determine the three-dimensional structure of the proliferating cell nuclear antigen (PCNA) in complex with a domain from the DNA polymerase Rev1. Rev1, a member of the translesion synthesis DNA polymerases, allows replication past DNA lesions, making it a key factor in DNA damage-induced mutagenesis in eukaryotes. Recent studies have shown that the high processivity of Rev1 at DNA lesions necessitates its interaction with PCNA. It was also shown that this interaction is stabilized during the DNA damage response. In agreement with these studies, our in vitro binding assays using calorimetry and NMR spectroscopy have demonstrated a tight interaction between PCNA and a BRCT domain located at the N-terminus Rev1. We have obtained crystals of the PCNA/Rev1-BRCT complex that are suitable for structure determination. PCNA was enriched in selenomethionine to collect MAD or SAD data.