Forward Genomics of Damage Control: An Undiscovered Class of Cancer Genes

该项目的中心假设是，存在一大类高度保守的 调节所有活细胞中自发DNA损伤水平的基因/蛋白质，也是如此 基因组不稳定的近端上游效应/调节器。这些建议是为了 包括许多高度保守的必需基因，在人类中，这些基因有望构成 一类新的癌症基因，不同于两个主要公认的基因类别(上游)： 基因组-看门人和看门人基因。然而，这些“损伤控制”基因/蛋白质 由于两个限制，一直没有被发现和认识。首先，没有技术 直接检测自发性/内源性DNA损伤，特别是DNA断裂 活细胞。因此，影响内源性DNA损伤水平的因素大多没有 做了化验，但没有考虑。其次，许多损害控制基因很可能是 对生物体的生存至关重要，以及目前在模式生物体中的正向遗传方法， 否则可能会发现其中的一些基因，是对必要基因的偏见。 我们预计损伤控制基因至少包括以下几个方面：(1)正常代谢 途径(碳代谢等)不可避免地产生有毒的副产品，与 造成碱基/核苷酸损伤的DNA；(2)清除这些副产物的蛋白质/分子； (3)DNA复制组件。(DNA断裂通常是复制造成损伤的结果。)所有的 它们被认为是非常保守的，因为它们是生命的基础，而且大多数是 被认为是必需的基因。 这个项目的目标是--(1)发展一种新的方法论范式 基因组学，将允许快速识别基因，包括必要的基因， 生物学中的任何问题；(2)使用这个范例，在简单的 模拟生物体大肠杆菌，鉴定它们在人类中的对应物，然后鉴定和验证 人类癌症候选基因。这些结果可能会形成理解什么是 被预测为第三类，非常大，高度保守，可能是重要的促癌类 突变。

The central hypothesis of this project is that there exists a large, highly-conserved class of genes/proteins that regulate levels of spontaneous DNA damage in all living cells, and so are the proximal, upstream effectors/modulators of genome instability. These are proposed to include many highly conserved essential genes that, in humans, are expected to constitute a ¿new¿ class of cancer genes distinct from (upstream of) the two main recognized classes: genomic-caretaker and gatekeeper genes. However these ¿damage-control¿ genes/proteins have remained undiscovered and unrecognized because of two limitations. First, no technology existed to assay spontaneous/endogenous DNA damage, particularly DNA breakage, directly in living cells. Thus, factors that affect the levels of endogenous DNA damage have mostly not been assayed, and not considered. Second, many of the damage-control genes are likely to be essential for organism viability, and current forward-genetic approaches in model organisms, which might otherwise have found some of these genes, are biased against essential genes. We expect the damage-control genes to encompass at least the following: (1) normal metabolic pathways (carbon metabolism, etc.) that unavoidably produce toxic by-products that react with DNA causing base/nucleotide damage; (2) proteins/molecules that scavenge these by-products; (3) DNA replication components. (DNA breaks often result from replication into damage.) All of these are expected to be very highly conserved because they are so basic to life, and most are expected to be essential genes. The goals of this project are--(1) to develop a new methodological paradigm of forward genomics that will allow rapid identification of genes, including essential genes, of interest to any problem in biology; (2) using this paradigm, to find the damage-control genes in the simple model organism E. coli, identify their counterparts in human, then identify and validate the candidate cancer genes in human. The results may form the basis of understanding what is predicted to be a third, very large, highly-conserved and potentially important class of cancerpromoting mutations.