Brumby Anthony M, Zraly Claudia B, Horsfield Julie A, Secombe Julie, Saint Robert, Dingwall Andrew K, Richardson Helena
Peter MacCallum Cancer Institute, Locked bag 1, A'Beckett Street, Melbourne, Victoria 8006, Australia.
EMBO J. 2002 Jul 1;21(13):3377-89. doi: 10.1093/emboj/cdf334.
Cyclin E-Cdk2 is essential for S phase entry. To identify genes interacting with cyclin E, we carried out a genetic screen using a hypomorphic mutation of Drosophila cyclin E (DmcycE(JP)), which gives rise to adults with a rough eye phenotype. Amongst the dominant suppressors of DmcycE(JP), we identified brahma (brm) and moira (mor), which encode conserved core components of the Drosophila Brm complex that is highly related to the SWI-SNF ATP-dependent chromatin remodeling complex. Mutations in genes encoding other Brm complex components, including snr1 (BAP45), osa and deficiencies that remove BAP60 and BAP111 can also suppress the DmcycE(JP) eye phenotype. We show that Brm complex mutants suppress the DmcycE(JP) phenotype by increasing S phases without affecting DmcycE protein levels and that DmcycE physically interacts with Brm and Snr1 in vivo. These data suggest that the Brm complex inhibits S phase entry by acting downstream of DmcycE protein accumulation. The Brm complex also physically interacts weakly with Drosophila retinoblastoma (Rbf1), but no genetic interactions were detected, suggesting that the Brm complex and Rbf1 act largely independently to mediate G(1) arrest.
细胞周期蛋白E - Cdk2对于进入S期至关重要。为了鉴定与细胞周期蛋白E相互作用的基因,我们利用果蝇细胞周期蛋白E(DmcycE(JP))的一个亚效突变进行了遗传筛选,该突变会导致成虫出现粗糙眼表型。在DmcycE(JP)的显性抑制因子中,我们鉴定出了brahma(brm)和moira(mor),它们编码果蝇Brm复合物的保守核心成分,该复合物与SWI - SNF ATP依赖性染色质重塑复合物高度相关。编码其他Brm复合物成分的基因突变,包括snr1(BAP45)、osa以及去除BAP60和BAP111的缺失突变,也能抑制DmcycE(JP)的眼表型。我们发现Brm复合物突变体通过增加S期来抑制DmcycE(JP)表型,而不影响DmcycE蛋白水平,并且DmcycE在体内与Brm和Snr1发生物理相互作用。这些数据表明,Brm复合物通过在DmcycE蛋白积累的下游发挥作用来抑制S期进入。Brm复合物也与果蝇视网膜母细胞瘤(Rbf1)发生微弱的物理相互作用,但未检测到遗传相互作用,这表明Brm复合物和Rbf1在很大程度上独立发挥作用来介导G(1)期阻滞。