Liu N, Lucibello F C, Zwicker J, Engeland K, Müller R
Institut für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität Marburg, Germany.
Nucleic Acids Res. 1996 Aug 1;24(15):2905-10. doi: 10.1093/nar/24.15.2905.
B-myb belongs to a group of cell cycle genes whose transcription is repressed in G0/early G1 through a binding site for the transcription factor E2F. Here, we show that the B-myb repressor element is specifically recognised by heterodimers consisting of DP-1 and E2F-1, E2F-3 or E2F-4. Surprisingly, E2F-mediated repression is dependent on a contiguous corepressor element that resembles the CHR previously established as a corepressor of the CDE in cell cycle genes derepressed in S/G2, such as cyclin A, cdc2 and cdc25C. A factor binding to the B-myb CHR was identified in fractionated HeLa nuclear extract and found to interact with the minor groove, as previously shown by in vivo footprinting for the cyclin A CHR. The B-myb and cdc25C CHRs are related with respect to protein binding but are functionally clearly distinct. Our results support a model where both E2F- and CDE-mediated repression, acting at different stages in the cell cycle, are dependent on promoter-specific CHR elements.
B-myb属于一组细胞周期基因,其转录在G0/早期G1期通过转录因子E2F的结合位点受到抑制。在此,我们表明B-myb抑制元件被由DP-1和E2F-1、E2F-3或E2F-4组成的异二聚体特异性识别。令人惊讶的是,E2F介导的抑制依赖于一个连续的共抑制元件,该元件类似于先前确定的作为细胞周期基因中CDE共抑制因子的CHR,这些基因在S/G2期去抑制,如细胞周期蛋白A、cdc2和cdc25C。在分级分离的HeLa细胞核提取物中鉴定出一种与B-myb CHR结合的因子,发现它与小沟相互作用,正如先前通过细胞周期蛋白A CHR的体内足迹分析所显示的那样。B-myb和cdc25C CHR在蛋白质结合方面相关,但在功能上明显不同。我们的结果支持一种模型,其中E2F和CDE介导的抑制作用于细胞周期的不同阶段,都依赖于启动子特异性的CHR元件。