Institut de Chimie Moléculaire, ICMUB CNRS UMR6302, UBFC, 21078 Dijon, France.
Institut Curie, PSL Research University, CNRS UMR9187, INSERM U1196, 91405 Orsay, France.
Nucleic Acids Res. 2018 Feb 16;46(3):e16. doi: 10.1093/nar/gkx1118.
The quest for chemicals able to operate at selected genomic loci in a spatiotemporally controlled manner is desirable to create manageable DNA damages. Mounting evidence now shows that alternative DNA structures, including G-quadruplexes and branched DNA (or DNA junctions), might hamper proper progression of replication fork, thus triggering DNA damages and genomic instability. Therefore, small molecules that stabilize these DNA structures are currently scrutinized as a promising way to create genomic defects that cannot be dealt with properly by cancer cells. While much emphasis has been recently given to G-quadruplexes and related ligands, we report herein on three-way DNA junctions (TWJ) and related ligands. We first highlight the biological implications of TWJ and their strategic relevance as triggers for replicative stress. Then, we describe a new in vitro high-throughput screening assay, TWJ-Screen, which allows for identifying TWJ ligands with both high affinity and selectivity for TWJ over other DNA structures (duplexes and quadruplexes), in a convenient and unbiased manner as demonstrated by the screening of a library of 25 compounds from different chemical families. TWJ-Screen thus represents a reliable mean to uncover molecular tools able to foster replicative stress through an innovative approach, thus providing new strategic opportunities to combat cancers.
人们希望寻找能够在特定基因组位置上以时空可控方式发挥作用的化学物质，以制造可管理的 DNA 损伤。越来越多的证据表明，包括 G-四链体和分支 DNA（或 DNA 连接点）在内的替代 DNA 结构可能会阻碍复制叉的正常前进，从而引发 DNA 损伤和基因组不稳定性。因此，目前正在研究能够稳定这些 DNA 结构的小分子，作为一种有前途的方法来制造无法被癌细胞正确处理的基因组缺陷。尽管最近人们非常关注 G-四链体和相关配体，但我们在此报告了三链 DNA 连接点（TWJ）及其相关配体。我们首先强调了 TWJ 的生物学意义及其作为复制应激触发器的战略相关性。然后，我们描述了一种新的体外高通量筛选测定法 TWJ-Screen，该方法可以方便且无偏地识别对 TWJ 具有高亲和力和选择性的 TWJ 配体，优于其他 DNA 结构（双链体和四链体），这一点通过对来自不同化学家族的 25 种化合物文库的筛选得到了证明。TWJ-Screen 因此代表了一种可靠的方法，可以通过创新的方法发现能够促进复制应激的分子工具，从而为对抗癌症提供新的战略机会。
