Studying essential DNA metabolism proteins in Xenopus egg extract.

V. Sannino, A. M. Kolinjivadi, G. Baldi, V. Costanzo
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引用次数: 8

Abstract

The correct duplication of genetic information is essential to maintain genome stability, which is lost in cancer cells. Replication fork integrity is ensured by a number of DNA metabolism proteins that assist replication of chromatin regions difficult to replicate due to their intrinsic DNA sequence composition, coordinate repair of DNA molecules resulting from aberrant replication events or protect replication forks in the presence of lesions impairing their progression. Some DNA metabolism genes involved in DNA repair are essential in higher eukaryotes even in unchallenged conditions, suggesting the existence of biological processes requiring these specialized functions in organisms with complex genomes. The impact on cell survival of null mutants of many DNA metabolism genes has precluded complete in depth analysis of their function. Cell free extracts represent a fundamental tool to overcome survival issues. The Xenopus laevis egg cell free extract is an ideal system to study replication-associated functions of essential genes. We are taking advantage of this system together with innovative imaging and proteomic based experimental approaches to characterize the molecular function of essential DNA metabolism proteins. Using this approach we have uncovered the role of some essential homologous recombination and fork protection proteins in chromosomal DNA replication and we have characterized some of the factors required for faithful replication of specific vertebrate genomic regions. This approach will be instrumental to study the molecular mechanisms underlying the function of a number of essential DNA metabolism proteins involved in the maintenance of genome stability in complex genomes.
非洲爪蟾卵提取物中必需DNA代谢蛋白的研究。
基因信息的正确复制对于维持基因组的稳定性至关重要,而这种稳定性在癌细胞中是缺失的。复制叉的完整性是由一些DNA代谢蛋白保证的,这些DNA代谢蛋白协助染色质区域的复制,这些染色质区域由于其固有的DNA序列组成而难以复制,协调由异常复制事件引起的DNA分子的修复,或者在存在损害其进展的病变时保护复制叉。一些参与DNA修复的DNA代谢基因在高等真核生物中是必不可少的,即使在没有挑战的条件下,这表明在具有复杂基因组的生物体中存在需要这些特殊功能的生物过程。许多DNA代谢基因的零突变体对细胞存活的影响阻碍了对其功能的完整深入分析。无细胞提取物是克服生存问题的基本工具。非洲爪蟾卵细胞提取液是研究重要基因复制相关功能的理想系统。我们正在利用该系统与创新的成像和基于蛋白质组学的实验方法来表征必需DNA代谢蛋白的分子功能。利用这种方法,我们已经发现了一些必要的同源重组和叉保护蛋白在染色体DNA复制中的作用,我们已经表征了特定脊椎动物基因组区域忠实复制所需的一些因素。这种方法将有助于研究一些基本DNA代谢蛋白在复杂基因组中参与维持基因组稳定性的功能的分子机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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