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Xenopus egg extract to study regulation of genome‐wide and locus‐specific DNA replication
Author(s) -
Raspelli Erica,
Falbo Lucia,
Costanzo Vincenzo
Publication year - 2017
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22996
Subject(s) - biology , xenopus , dna replication , eukaryotic dna replication , control of chromosome duplication , origin recognition complex , dna repair , dna re replication , genome , genetics , pre replication complex , gene , dna replication factor cdt1 , gene duplication , dna damage , dna , microbiology and biotechnology , replication protein a , chromatin , replication timing , dna binding protein , transcription factor
Faithful DNA replication, coupled with accurate repair of DNA damage, is essential to maintain genome stability and relies on different DNA metabolism genes. Many of these genes are involved in the assembly of replication origins, in the coordination of DNA repair to protect replication forks progression in the presence of DNA damage and in the replication of repetitive chromatin regions. Some DNA metabolism genes are essential in higher eukaryotes, suggesting the existence of specialized mechanisms of repair and replication in organisms with complex genomes. The impact on cell survival of many of these genes has so far precluded in depth molecular analysis of their function. The cell‐free Xenopus laevis egg extract represents an ideal system to overcome survival issues and to facilitate the biochemical study of replication‐associated functions of essential proteins in vertebrate organisms. Here, we will discuss how Xenopus egg extracts have been used to study cellular and molecular processes, such as DNA replication and DNA repair. In particular, we will focus on innovative imaging and proteomic‐based experimental approaches to characterize the molecular function of a number of essential DNA metabolism factors involved in the duplication of complex vertebrate genomes.