Udayakumar T S, Belakavadi Madesh, Choi Kyoung-Han, Pandey Pradeep K, Fondell Joseph D
Department of Physiology & Biophysics, Robert Wood Johnson Medical School, UMDNJ, Piscataway, NJ 08854, USA.
J Biol Chem. 2006 May 26;281(21):14691-9. doi: 10.1074/jbc.M600163200. Epub 2006 Mar 30.
The TRAP/Mediator coactivator complex serves as a functional interface between DNA-bound transactivators and the RNA polymerase II-associated basal transcription apparatus. TRAP220/MED1 is a variably associated subunit of the complex that plays a specialized role in selectively targeting TRAP/Mediator to specific genes. Ablation of the Trap220/Med1 gene in mice impairs embryonic cell growth, yet the underlying mechanism is unknown. In this report, we identified distinct cell growth regulatory genes whose expression is affected by the loss of TRAP220/MED1 by RNA interference. Among the down-regulated genes revealed by cDNA microarray analyses, we identified Aurora-A, a centrosome kinase that plays a critical role in regulating M phase events and is frequently amplified in several types of cancer. In general, we found that TRAP220/MED1 expression is required for high basal levels of Aurora-A gene expression and that ectopic overexpression of TRAP220/MED1 coactivates transcription from the Aurora-A gene promoter. Furthermore, chromatin immunoprecipitation assays show that TRAP220/MED1-containing TRAP/Mediator complexes directly bind to the Aurora-A promoter in vivo. Finally, we present evidence suggesting that TRAP/Mediator is recruited to the Aurora-A gene via direct interactions between TRAP220/MED1 and the Ets-related transcription factor GABP. Taken together, these findings suggest that TRAP220/MED1 plays a novel coregulatory role in facilitating the recruitment of TRAP/Mediator to specific target genes involved in growth and cell cycle progression.
TRAP/中介体共激活复合物充当DNA结合反式激活因子与RNA聚合酶II相关基础转录装置之间的功能接口。TRAP220/MED1是该复合物中一个可变关联的亚基,在将TRAP/中介体选择性靶向特定基因方面发挥着特殊作用。在小鼠中敲除Trap220/Med1基因会损害胚胎细胞生长,但其潜在机制尚不清楚。在本报告中,我们通过RNA干扰鉴定了不同的细胞生长调节基因,其表达受TRAP220/MED1缺失的影响。在cDNA微阵列分析揭示的下调基因中,我们鉴定出Aurora-A,一种中心体激酶,它在调节M期事件中起关键作用,并且在几种类型的癌症中经常扩增。一般来说,我们发现TRAP220/MED1的表达是Aurora-A基因高基础表达水平所必需的,并且TRAP220/MED1的异位过表达可共激活Aurora-A基因启动子的转录。此外,染色质免疫沉淀试验表明,含有TRAP220/MED1的TRAP/中介体复合物在体内直接结合到Aurora-A启动子上。最后,我们提供的证据表明,TRAP/中介体通过TRAP220/MED1与Ets相关转录因子GABP之间的直接相互作用被招募到Aurora-A基因。综上所述,这些发现表明TRAP220/MED1在促进TRAP/中介体招募到参与生长和细胞周期进程的特定靶基因方面发挥着新的共调节作用。