Brodsky Michael H, Weinert Brian T, Tsang Garson, Rong Yikang S, McGinnis Nadine M, Golic Kent G, Rio Donald C, Rubin Gerald M
Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Mol Cell Biol. 2004 Feb;24(3):1219-31. doi: 10.1128/MCB.24.3.1219-1231.2004.
We have used genetic and microarray analysis to determine how ionizing radiation (IR) induces p53-dependent transcription and apoptosis in Drosophila melanogaster. IR induces MNK/Chk2-dependent phosphorylation of p53 without changing p53 protein levels, indicating that p53 activity can be regulated without an Mdm2-like activity. In a genome-wide analysis of IR-induced transcription in wild-type and mutant embryos, all IR-induced increases in transcript levels required both p53 and the Drosophila Chk2 homolog MNK. Proapoptotic targets of p53 include hid, reaper, sickle, and the tumor necrosis factor family member EIGER: Overexpression of Eiger is sufficient to induce apoptosis, but mutations in Eiger do not block IR-induced apoptosis. Animals heterozygous for deletions that span the reaper, sickle, and hid genes exhibited reduced IR-dependent apoptosis, indicating that this gene complex is haploinsufficient for induction of apoptosis. Among the genes in this region, hid plays a central, dosage-sensitive role in IR-induced apoptosis. p53 and MNK/Chk2 also regulate DNA repair genes, including two components of the nonhomologous end-joining repair pathway, Ku70 and Ku80. Our results indicate that MNK/Chk2-dependent modification of Drosophila p53 activates a global transcriptional response to DNA damage that induces error-prone DNA repair as well as intrinsic and extrinsic apoptosis pathways.
我们利用基因和微阵列分析来确定电离辐射(IR)如何在黑腹果蝇中诱导p53依赖性转录和凋亡。IR诱导p53的MNK/Chk2依赖性磷酸化,而不改变p53蛋白水平,这表明p53活性可以在没有类似Mdm2活性的情况下受到调节。在对野生型和突变型胚胎中IR诱导转录的全基因组分析中,所有IR诱导的转录水平增加都需要p53和果蝇Chk2同源物MNK。p53的促凋亡靶标包括hid、reaper、sickle以及肿瘤坏死因子家族成员EIGER:Eiger的过表达足以诱导凋亡,但Eiger的突变并不阻断IR诱导的凋亡。跨越reaper、sickle和hid基因的缺失杂合动物表现出IR依赖性凋亡减少,表明该基因复合体在诱导凋亡方面单倍剂量不足。在该区域的基因中,hid在IR诱导的凋亡中起核心的、剂量敏感的作用。p53和MNK/Chk2还调节DNA修复基因,包括非同源末端连接修复途径的两个组分Ku70和Ku80。我们的结果表明,果蝇p53的MNK/Chk2依赖性修饰激活了对DNA损伤的全局转录反应,该反应诱导易出错的DNA修复以及内在和外在凋亡途径。