a Polish Academy of Sciences , Institute of Bioorganic Chemistry , Poznan , Poland.
RNA Biol. 2019 Jan;16(1):25-41. doi: 10.1080/15476286.2018.1556084. Epub 2018 Dec 20.
Transcription initiation sites of Trp53 gene in mice were determined using the 5'RACE method. Based on sequence alignment of the 5'-terminal regions of p53 mRNA in mammals, the site for the most abundant transcript turned out to be essentially identical with that determined for human TP53 gene and slightly differed for the longest transcripts, in mice and humans. Secondary structures of the 5' -terminal regions of the shorter, most abundant and the longest mouse transcripts were determined in vitro and the shorter transcript was also mapped in transfected mouse cells. For the first time, secondary structure models of the 5' terminus of two mouse p53 mRNAs were proposed. Comparing these models with the conservativeness of the nucleotide sequence of the 5'-terminal region of mRNA in mouse and other mammals, the possible function of the selected structural domains of this region was discussed. To elucidate the translation mechanisms, the two studied mRNAs were translated in the presence of an increasing concentration of the cap analog. For the longest transcript, the data suggested that IRES element(s) was/were involved in translation initiation. Additionally, changes in p53 synthesis under genotoxic and endoplasmic reticulum stress conditions in mouse cells were analyzed.
采用 5'RACE 方法确定小鼠 Trp53 基因的转录起始位点。基于哺乳动物 p53 mRNA 5' 端区域的序列比对,最丰富的转录本的位点与人类 TP53 基因确定的位点基本相同,而在小鼠和人类中最长的转录本略有不同。体外测定了较短、最丰富和最长的小鼠转录本 5' 端的二级结构,并在转染的小鼠细胞中对较短的转录本进行了作图。首次提出了两种小鼠 p53 mRNA 5' 端的二级结构模型。将这些模型与小鼠和其他哺乳动物 mRNA 5' 端序列的保守性进行比较,讨论了该区域选定结构域的可能功能。为了阐明翻译机制,在存在帽类似物浓度增加的情况下,对这两种研究的 mRNA 进行了翻译。对于最长的转录本,数据表明 IRES 元件(s)参与了翻译起始。此外,还分析了小鼠细胞在遗传毒性和内质网应激条件下 p53 合成的变化。