Institute of Life Sciences, Hyderabad, Andhra Pradesh 500 046, India.
J Cell Biochem. 2013 Apr;114(4):899-907. doi: 10.1002/jcb.24429.
p53 protein, the central molecule of the apoptosis pathway, is mutated in 50% of the human cancers. Of late, p53 homologues have been identified from different invertebrates including Drosophila melanogaster, Caenorhabditis elegans, Squid, and Clams. We report the identification of a p53-like protein in Spodoptera frugiperda (Sf9) insect cells, which is activated during oxidative stress, caused by exposure to UV-B or H(2) O(2) , and binds to p53 consensus DNA binding motifs as well as other p53 cognate motifs. Sf9 p53 motif-binding protein is similar to murine and Drosophila p53 in terms of molecular size, which is around 50-60 kDa, as evident from UV cross-linking, and displays DNA binding characteristics similar to both insect and vertebrate p53 as seen from electrophoretic mobility shift assays. The N-terminal sequencing of the purified Sf9 p53 motif-binding protein reveals extensive homology to the pro-apoptotic FK-506 binding protein (FKBP-46), earlier identified in Sf9 cells as a factor which interacts with murine casein kinase. FKBP, an evolutionarily conserved protein of mammalian origin functions as a pro-apoptotic factor. Identification of FKBP-46 as a novel p53 motif-binding protein in insect cells adds a new facet to our understanding of the mechanisms of apoptosis under oxidative stress in the absence of a typical p53 homologue.
p53 蛋白是细胞凋亡途径的核心分子,在 50%的人类肿瘤中发生突变。最近,已经从不同的无脊椎动物中鉴定出 p53 同源物,包括果蝇、秀丽隐杆线虫、鱿鱼和蛤蜊。我们报告了在 Spodoptera frugiperda(Sf9)昆虫细胞中鉴定出一种 p53 样蛋白,该蛋白在暴露于 UV-B 或 H2O2 引起的氧化应激中被激活,并与 p53 共有 DNA 结合基序以及其他 p53 同源基序结合。Sf9 p53 基序结合蛋白在分子大小方面与鼠和果蝇 p53 相似,大约为 50-60 kDa,这从 UV 交联实验中可以明显看出,并从电泳迁移率变动分析中显示出与昆虫和脊椎动物 p53 相似的 DNA 结合特性。纯化的 Sf9 p53 基序结合蛋白的 N 端测序显示与促凋亡 FK-506 结合蛋白(FKBP-46)有广泛的同源性,FKBP-46 先前在 Sf9 细胞中被鉴定为与鼠酪蛋白激酶相互作用的因子。FKBP 是一种来源于哺乳动物的进化上保守的蛋白质,作为一种促凋亡因子发挥作用。鉴定 FKBP-46 为昆虫细胞中新型 p53 基序结合蛋白,为我们在没有典型 p53 同源物的情况下理解氧化应激下细胞凋亡的机制增添了新的内容。