Hochheimer Andreas, Zhou Sharleen, Zheng Shuang, Holmes Michael C, Tjian Robert
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley 94720-3204, USA.
Nature. 2002 Nov 28;420(6914):439-45. doi: 10.1038/nature01167.
Drosophila TATA-box-binding protein (TBP)-related factor 2 (TRF2) is a member of a family of TBP-related factors present in metazoan organisms. Recent evidence suggests that TRF2s are required for proper embryonic development and differentiation. However, true target promoters and the mechanisms by which TRF2 operates to control transcription remain elusive. Here we report the antibody affinity purification of a Drosophila TRF2-containing complex that contains components of the nucleosome remodelling factor (NURF) chromatin remodelling complex as well as the DNA replication-related element (DRE)-binding factor DREF. This latter finding led us to potential target genes containing TRF2-responsive promoters. We have used a combination of in vitro and in vivo assays to show that the DREF-containing TRF2 complex directs core promoter recognition of the proliferating cell nuclear antigen (PCNA) gene. We also identified additional TRF2-responsive target genes involved in DNA replication and cell proliferation. These data suggest that TRF2 functions as a core promoter-selectivity factor responsible for coordinating transcription of a subset of genes in Drosophila.
果蝇TATA框结合蛋白（TBP）相关因子2（TRF2）是后生动物中存在的TBP相关因子家族的成员。最近的证据表明，TRF2对于正常的胚胎发育和分化是必需的。然而，真正的靶启动子以及TRF2控制转录的机制仍然不清楚。在这里，我们报告了一种含果蝇TRF2的复合物的抗体亲和纯化，该复合物包含核小体重塑因子（NURF）染色质重塑复合物的成分以及DNA复制相关元件（DRE）结合因子DREF。后一个发现使我们找到了含有TRF2反应性启动子的潜在靶基因。我们使用了体外和体内试验相结合的方法，表明含DREF的TRF2复合物指导增殖细胞核抗原（PCNA）基因的核心启动子识别。我们还鉴定了参与DNA复制和细胞增殖的其他TRF2反应性靶基因。这些数据表明，TRF2作为一种核心启动子选择性因子，负责协调果蝇中一部分基因的转录。
