E2F integrates cell cycle progression with DNA repair, replication, and G2/M checkpoints
E2F integrates cell cycle progression with DNA repair, replication, and G2/M checkpoints
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DOI：
10.1101/gad.949802
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发表时间：
2002-01-15
影响因子：
10.5
通讯作者：
Dynlacht, BD
中科院分区：
文献类型：
--
作者：
Ren, B;Cam, H;Dynlacht, BD
已知E2F转录因子家族在细胞周期进程和增殖所需基因的及时表达中起关键作用，但仅鉴定出少数E2F靶基因。我们通过鉴定活的人类细胞中与E2F结合的其他基因，探索了E2F调节因子发挥更广泛作用的可能性。开发了一种方案，以确定哺乳动物细胞中DNA结合因子的基因组结合位点，该方案结合了交联蛋白-DNA复合物的免疫沉淀和DNA微阵列分析。在细胞周期进入过程中表达的大约1200个基因中，我们发现127个启动子在原代成纤维细胞中与E2F4转录因子结合。这些靶标的一个子集也被E2F1结合。大多数先前确定的靶基因已知在DNA复制和细胞周期控制中起作用，并在微阵列上表示，通过该分析得到了证实。我们还发现了一系列与E2F调控没有关联的基因，包括编码DNA损伤检查点和修复途径组分的基因，以及涉及染色质组装/凝聚、染色体分离和有丝分裂纺锤体检查点的因子。我们的数据表明，E2F直接将细胞周期进程与DNA合成及其监视所必需的基因的协调调节联系起来。
The E2F transcription factor family is known to play a key role in the timely expression of genes required for cell cycle progression and proliferation, but only a few E2F target genes have been identified. We explored the possibility that E2F regulators play a broader role by identifying additional genes bound by E2F in living human cells. A protocol was developed to identify genomic binding sites for DNA-binding factors in mammalian cells that combines immunoprecipitation of cross-linked protein-DNA complexes with DNA microarray analysis. Among similar to1200 genes expressed during cell cycle entry, we found that the promoters of 127 were bound by the E2F4 transcription factor in primary fibroblasts. A subset of these targets was also bound by E2F1. Most previously identified target genes known to have roles in DNA replication and cell cycle control and represented on the microarray were confirmed by this analysis. We also identified a remarkable cadre of genes with no previous connection to E2F regulation, including genes that encode components of the DNA damage checkpoint and repair pathways, as well as factors involved in chromatin assembly/ condensation, chromosome segregation, and the mitotic spindle checkpoint. Our data indicate that E2F directly links cell cycle progression with the coordinate regulation of genes essential for both the synthesis of DNA as well as its surveillance.