Fan Lin, Jiang Jiajun, Gao Jinning, Song Huayu, Liu Jinxiang, Yang Likun, Li Zan, Chen Yan, Zhang Quanqi, Wang Xubo
Key Laboratory of Marine Genetics and Breeding (Ocean University of China), Ministry of Education, Qingdao 266003, China.
Int J Mol Sci. 2015 Apr 23;16(5):9097-118. doi: 10.3390/ijms16059097.
PRDM14 is a PR (PRDI-BF1-RIZ1 homologous) domain protein with six zinc fingers and essential roles in genome-wide epigenetic reprogramming. This protein is required for the establishment of germ cells and the maintenance of the embryonic stem cell ground state. In this study, we cloned the full-length cDNA and genomic DNA of the Paralichthys olivaceus prdm14 (Po-prdm14) gene and isolated the 5' regulatory region of Po-prdm14 by whole-genome sequencing. Peptide sequence alignment, gene structure analysis, and phylogenetic analysis revealed that Po-PRDM14 was homologous to mammalian PRDM14. Results of real-time quantitative polymerase chain reaction amplification (RT-qPCR) and in situ hybridization (ISH) in embryos demonstrated that Po-prdm14 was highly expressed between the morula and late gastrula stages, with its expression peaking in the early gastrula stage. Relatively low expression of Po-prdm14 was observed in the other developmental stages. ISH of gonadal tissues revealed that the transcripts were located in the nucleus of the oocytes in the ovaries but only in the spermatogonia and not the spermatocytes in the testes. We also presume that the Po-prdm14 transcription factor binding sites and their conserved binding region among vertebrates. The combined results suggest that Po-PRDM14 has a conserved function in teleosts and mammals.
PRDM14是一种具有六个锌指结构的PR(PRDI-BF1-RIZ1同源)结构域蛋白,在全基因组表观遗传重编程中发挥着重要作用。该蛋白对于生殖细胞的建立和胚胎干细胞基态的维持是必需的。在本研究中,我们克隆了褐牙鲆prdm14(Po-prdm14)基因的全长cDNA和基因组DNA,并通过全基因组测序分离出Po-prdm14的5'调控区。肽序列比对、基因结构分析和系统发育分析表明,Po-PRDM14与哺乳动物PRDM14同源。胚胎中的实时定量聚合酶链反应扩增(RT-qPCR)和原位杂交(ISH)结果表明,Po-prdm14在桑椹胚和原肠胚晚期之间高表达,在原肠胚早期表达达到峰值。在其他发育阶段观察到Po-prdm14表达相对较低。性腺组织的ISH显示,转录本位于卵巢卵母细胞的细胞核中,但仅位于睾丸的精原细胞中,而不在精母细胞中。我们还推测了Po-prdm14转录因子结合位点及其在脊椎动物中的保守结合区域。综合结果表明,Po-PRDM14在硬骨鱼和哺乳动物中具有保守功能。