Division of Animal Sciences, Agharkar Research Institute, Pune, India.
PLoS One. 2013;8(4):e61062. doi: 10.1371/journal.pone.0061062. Epub 2013 Apr 8.
Hydra, one of the earliest metazoans with tissue grade organization and nervous system, is an animal with a remarkable regeneration capacity and shows no signs of organismal aging. We have for the first time identified genes of the nucleotide excision repair (NER) pathway from hydra. Here we report cloning and characterization of hydra homolog of xeroderma pigmentosum group F (XPF) gene that encodes a structure-specific 5' endonuclease which is a crucial component of NER. In silico analysis shows that hydra XPF amino acid sequence is very similar to its counterparts from other animals, especially vertebrates, and shows all features essential for its function. By in situ hybridization, we show that hydra XPF is expressed prominently in the multipotent stem cell niche in the central region of the body column. Ectoderm of the diploblastic hydra was shown to express higher levels of XPF as compared to the endoderm by semi-quantitative RT-PCR. Semi-quantitative RT-PCR analysis also demonstrated that interstitial cells, a multipotent and rapidly cycling stem cell lineage of hydra, express higher levels of XPF mRNA than other cell types. Our data show that XPF and by extension, the NER pathway is highly conserved during evolution. The prominent expression of an NER gene in interstitial cells may have implications for the lack of senescence in hydra.
水螅是最早具有组织层次和神经系统的后生动物之一,是一种具有显著再生能力且没有明显衰老迹象的动物。我们首次从水螅中鉴定出核苷酸切除修复(NER)途径的基因。在这里,我们报告了水螅 Xeroderma Pigmentosum 组 F(XPF)基因同源物的克隆和特征,该基因编码一种结构特异性的 5'内切核酸酶,是 NER 的关键组成部分。计算机分析表明,水螅 XPF 氨基酸序列与来自其他动物,尤其是脊椎动物的同源物非常相似,并且具有其功能所必需的所有特征。通过原位杂交,我们发现水螅 XPF 在身体柱中央的多能干细胞龛中表达显著。通过半定量 RT-PCR 显示,与内胚层相比,二胚层水螅的外胚层表达更高水平的 XPF。半定量 RT-PCR 分析还表明,水螅的间质细胞(一种多能且快速分裂的干细胞谱系)表达更高水平的 XPF mRNA 比其他细胞类型。我们的数据表明,XPF 及其扩展的 NER 途径在进化过程中高度保守。NER 基因在间质细胞中的显著表达可能与水螅缺乏衰老有关。