Jackman J, Alamo I, Fornace A J
Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, Maryland 20892.
Cancer Res. 1994 Nov 1;54(21):5656-62.
The growth arrest and DNA damage-inducible (gadd) genes represent a group of five stress-inducible genes that are coordinately regulated at the transcriptional level. Posttranscriptional regulation of gadd153, gadd45, gadd34, gadd33, and gadd7 was studied after exposure to DNA-damaging agents or other growth arrest treatments in hamster cells. Relative transcript levels were measured following treatment with the transcriptional inhibitor actinomycin D. After exposure to methylmethane sulfonate or UV radiation, all five gadd messages demonstrated a coordinate increase in mRNA stability compared to untreated exponentially growing cells. This enhanced stability was not an universal response to genotoxic stress since other DNA damage-inducible genes, such as c-jun and c-fos, did not show an appreciable increase in mRNA half-life. In contrast, induction of growth arrest by media depletion (starvation) or by treatment with the growth inhibitor prostaglandin A2 did not induce such an increase in mRNA stability in all gadd genes. Comparison of overall RNA turnover by 3H labeling of total cellular RNA also indicated that the preferential stabilization of the gadd transcripts by DNA-damaging agents was not an artifactual response due to variations in overall RNA metabolism within each treatment group. However, DNA-damaging agents were ineffective in inducing stabilization of gadd153 mRNA in growth-arrested cells. This suggest that the signal(s) that give rise to gadd mRNA stability may also be affected by the state of cellular proliferation. Together, these results suggest that the global posttranscriptional response of the gadd genes to DNA-damaging agents is specific and unique to actively growing cells, and further implicates the role of the gadd genes in the DNA damage response of cycling cells.
生长停滞与DNA损伤诱导(gadd)基因代表一组五个应激诱导基因,它们在转录水平上受到协同调控。在仓鼠细胞中,研究了gadd153、gadd45、gadd34、gadd33和gadd7在暴露于DNA损伤剂或其他生长停滞处理后的转录后调控。在用转录抑制剂放线菌素D处理后,测量相对转录水平。暴露于甲基磺酸甲酯或紫外线辐射后,与未处理的指数生长细胞相比,所有五个gadd信息在mRNA稳定性上均表现出协同增加。这种增强的稳定性并非对基因毒性应激的普遍反应,因为其他DNA损伤诱导基因,如c-jun和c-fos,在mRNA半衰期上并未显示出明显增加。相反,通过培养基耗尽(饥饿)或用生长抑制剂前列腺素A2处理诱导生长停滞,并未在所有gadd基因中诱导mRNA稳定性的这种增加。通过对总细胞RNA进行3H标记来比较总体RNA周转,也表明DNA损伤剂对gadd转录本的优先稳定作用并非由于每个处理组内总体RNA代谢的变化而产生的人为反应。然而,DNA损伤剂在生长停滞细胞中诱导gadd153 mRNA稳定化方面无效。这表明导致gadd mRNA稳定性的信号也可能受到细胞增殖状态的影响。总之,这些结果表明,gadd基因对DNA损伤剂的整体转录后反应对于活跃生长的细胞是特异且独特的,并进一步暗示了gadd基因在循环细胞的DNA损伤反应中的作用。