Madden S L, Galella E A, Zhu J, Bertelsen A H, Beaudry G A
Department of Molecular & Cellular Biology, PharmaGenics, Inc., Allendale, NJ 07401, USA.
Oncogene. 1997 Aug 28;15(9):1079-85. doi: 10.1038/sj.onc.1201091.
Serial analysis of gene expression (SAGE) allows for a quantitative, representative, and comprehensive profile of gene expression. We have utilized SAGE technology to contrast the differential gene expression profile in rat embryo fibroblast cells producing temperature-sensitive p53 tumor suppressor protein at permissive or non-permissive temperatures. Analysis of approximately 15,000 genes revealed that the expression of 14 genes (P < 0.001, > or = 0.03% abundance) was dependent on functional p53 protein, whereas the expression of three genes was significantly higher in cells producing non-functional p53 protein. Those genes whose expression was increased by functional p53 include RAS, U6 snRNA, cyclin G, EGR-1, and several novel genes. The expression of actin, tubulin, and HSP70 genes was elevated at the non-permissive temperature for p53 function. Interestingly, the expression of several genes was dependent on a non-temperature-sensitive mutant p53 suggesting altered transcription profiles dependent on specific p53 mutant proteins. These results demonstrate the utility of SAGE for rapidly and reproducibly evaluating global transcriptional responses within different cell populations.
基因表达系列分析(SAGE)能够实现对基因表达的定量、代表性和全面性描述。我们利用SAGE技术对比了在允许温度或非允许温度下产生温度敏感型p53肿瘤抑制蛋白的大鼠胚胎成纤维细胞中的差异基因表达谱。对大约15000个基因的分析表明,14个基因(P < 0.001,丰度>或= 0.03%)的表达依赖于功能性p53蛋白,而3个基因在产生非功能性p53蛋白的细胞中表达显著更高。那些表达因功能性p53而增加的基因包括RAS、U6 snRNA、细胞周期蛋白G、早期生长反应蛋白1(EGR-1)以及几个新基因。在p53功能的非允许温度下,肌动蛋白、微管蛋白和热休克蛋白70(HSP70)基因的表达升高。有趣的是,几个基因的表达依赖于非温度敏感型突变p53,这表明转录谱的改变依赖于特定的p53突变蛋白。这些结果证明了SAGE在快速且可重复地评估不同细胞群体内全局转录反应方面的实用性。