A network of multi-tasking proteins at the DNA replication fork preserves genome stability.
A network of multi-tasking proteins at the DNA replication fork preserves genome stability.
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DOI：
10.1371/journal.pgen.0010061
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发表时间：
2005-12
期刊：
影响因子：
4.5
通讯作者：
Campbell JL
中科院分区：
文献类型：
--
作者：
Budd ME;Tong AH;Polaczek P;Peng X;Boone C;Campbell JL
为了阐明维持高保真基因组复制的网络，我们将DNA复制基因DNA2的两个条件突变等位基因引入到大约4,700个活酵母缺失突变体中，并确定了双突变体的适合度。鉴定出56个dna - 2相互作用基因。对43个dna2相互作用基因中的每一个的基因组合成致死谱进行聚类分析，确定了一个网络（由322个基因和876个相互作用组成），其拓扑结构为复制蛋白如何协调调节和修复以保护基因组完整性提供了线索。这些结果还揭示了疑问基因DNA2的功能，尽管多年的研究仍然存在争议，特别是它在冈崎片段加工中的作用及其在体内底物的性质。由于复制叉上几乎所有蛋白质的多功能性质，任何单一基因相互作用的意义本质上都是模糊的。然而，当前研究的多路复用性质，结合后续的支持实验，揭示了大多数（如果不是全部的话）需要Dna2p的独特途径。这些不仅包括冈崎片段加工和DNA修复，还包括染色质动力学。一代又一代保持基因组的稳定性是防止突变和随之而来的疾病的主要防御手段。因此，细胞进化出了复杂的机制，包括冗余的、部分重叠的通路，以保护基因组遗传的保真度。利用现代基因筛选技术，研究人员可以研究酵母中可能参与这些途径的每个基因，他们已经定义了一个由322个基因组成的网络，这些基因共同保护DNA复制过程。以前的方法仅限于定义一个或几个基因的相互作用，但是影响所有非必需酵母基因的突变的可用性允许在本研究中确定超过800种相互作用。此外，本研究中使用的合成基因阵列技术允许鉴定酵母中与基本复制蛋白Dna2p相互作用的每个非必需基因。该方法的全面性，即使不是全部，也能在一次实验中识别出多任务Dna2p参与的大部分途径。与传统的低通量遗传方法相比，该研究的基因组规模显着加速了对这种蛋白质的理解。
To elucidate the network that maintains high fidelity genome replication, we have introduced two conditional mutant alleles of DNA2, an essential DNA replication gene, into each of the approximately 4,700 viable yeast deletion mutants and determined the fitness of the double mutants. Fifty-six DNA2-interacting genes were identified. Clustering analysis of genomic synthetic lethality profiles of each of 43 of the DNA2-interacting genes defines a network (consisting of 322 genes and 876 interactions) whose topology provides clues as to how replication proteins coordinate regulation and repair to protect genome integrity. The results also shed new light on the functions of the query gene DNA2, which, despite many years of study, remain controversial, especially its proposed role in Okazaki fragment processing and the nature of its in vivo substrates. Because of the multifunctional nature of virtually all proteins at the replication fork, the meaning of any single genetic interaction is inherently ambiguous. The multiplexing nature of the current studies, however, combined with follow-up supporting experiments, reveals most if not all of the unique pathways requiring Dna2p. These include not only Okazaki fragment processing and DNA repair but also chromatin dynamics. Maintenance of genome stability from generation to generation is a primary defense against mutation and ensuing disease. Thus, the cell has evolved complex mechanisms, consisting of redundant, partially overlapping pathways, to protect the fidelity of genome inheritance. Using modern genetic screening techniques that allow one to investigate every gene in yeast that might be involved in these pathways, the researchers have defined a network consisting of 322 genes that together safeguard the DNA replication process. Previous approaches were limited to defining the interaction of one or a few genes, but the availability of mutants affecting all of the nonessential yeast genes allowed the identification of over 800 interactions in this study. In addition, the synthetic genetic array technique used in this study allowed identification of every nonessential gene in yeast that interacts with an essential replication protein, Dna2p. The comprehensiveness of the approach identified most, if not all, of the pathways in which the multitasking Dna2p participates, in a single experiment. The genomic scale of the study significantly accelerates understanding of this protein over traditional, low-throughput genetic methods.
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影响因子：
5.3
作者：
Budd, ME;Campbell, JL
通讯作者：
Campbell, JL
影响因子：
10.5
作者：
BRACHMANN, CB;SHERMAN, JM;BOEKE, JD
通讯作者：
BOEKE, JD
影响因子：
64.8
作者：
Bae, SH;Bae, KH;Seo, YS
通讯作者：
Seo, YS
影响因子：
3.3
作者：
Fiorentino, DF;Crabtree, GR
通讯作者：
Crabtree, GR
DOI：
10.1016/s0921-8777(99)00072-5
发表时间：
2000-04-28
期刊：
MUTATION RESEARCH-DNA REPAIR
影响因子：
--
作者：
Budd, ME;Campbell, JL
通讯作者：
Campbell, JL