Conflicts between DNA replication fork progression and transcriptional regulation

所有生物都必须复制它们的遗传物质，这样才能代代相传。然而，这一过程发生在拥挤的环境中，因为其他同样重要的细胞功能必须与DNA复制同时发生。因此，即使DNA上覆盖着许多DNA结合蛋白，也必须发生基因组复制。人们对这些DNA结合蛋白对复制的影响知之甚少，尽管有很多间接证据表明这种碰撞会导致DNA突变。我们的目标是使用细菌的复制机制来分析与DNA结合的蛋白质对复制的影响。我们将测量DNA结合蛋白停止复制机制的可能性，并将表征导致复制受阻的蛋白质-DNA复合体的性质。然而，细胞显然有能力复制它们的基因组，即使DNA被一层蛋白质包裹着。因此，我们将寻找能够帮助复制机制在DNA-蛋白质复合体中移动的酶。这些实验将帮助我们了解所有细胞如何在复杂的富含蛋白质的环境中复制它们的基因组。它们还可能揭示细胞如何将其复制机制停滞并导致潜在有害突变的可能性降至最低。

All organisms must duplicate their genetic material so that it can be passed from one generation to the next. However, this process occurs in a crowded environment since other equally important cellular functions must occur at the same time as DNA replication. As a result, genome duplication must occur even though the DNA is coated with many DNA binding proteins. Little is known of the effects of these DNA binding proteins upon replication even though there is much indirect evidence that such collisions lead to mutations in the DNA. We aim to use the replication machinery from a bacterium to analyse the effects of proteins bound to the DNA upon replication. We will measure the probability of DNA binding proteins halting the replication machinery, and will characterise the properties of protein-DNA complexes which lead to blockage of replication. However, cells clearly do have the ability to replicate their genomes even though the DNA is coated in a panoply of proteins. We will therefore search for enzymes that can help the replication machinery to move through DNA-protein complexes. These experiments will help us understand how all cells duplicate their genomes in a complex protein-rich environment. They may also cast light on how cells minimise the chances of their replication machinery stalling and causing potentially harmful mutations.