Mokdad-Gargouri R, Belhadj K, Gargouri A
Laboratoire 'Génétique Moléculaire des Eucaryotes', Centre de Biotechnologie de Sfax, BP'K'3038, Sfax-Tunisia.
Nucleic Acids Res. 2001 Mar 1;29(5):1222-7. doi: 10.1093/nar/29.5.1222.
We have expressed human p53 cDNA in the yeast Saccharomyces cerevisiae and shown that the level of production and the length of the p53 protein depends on the presence of untranslated mRNA regions (UTRs). The expression of the ORF alone leads to a p53 protein of correct size (53 kDa) that accumulates to high levels, concomitantly with the presence of a small amount of a p40 protein (40 kDa). However, when either the entire 5'-UTR and a part of the 3'- or 5'-UTR alone is used, this leads to the production of small amounts of the 40 kDa truncated form only. The p40 protein corresponds to a truncated form of p53 at the C-terminal extremity since it reacts only with a monoclonal antibody recognising the N-terminal epitope. This effect on the amount and length of p53 protein had no correlation at the mRNA level, suggesting that translational control probably occurs through the 5'-UTR. We propose a model of structural interaction between this UTR and a part of the ORF mRNA for the regulation of p53 expression in this heterologous context.
我们已在酿酒酵母中表达了人p53 cDNA，并表明p53蛋白的产生水平和长度取决于非翻译mRNA区域（UTR）的存在。仅开放阅读框（ORF）的表达会产生大小正确（53 kDa）的p53蛋白，其会积累到高水平，同时存在少量p40蛋白（40 kDa）。然而，当单独使用整个5'-UTR以及部分3'-或5'-UTR时，这只会导致少量40 kDa截短形式的产生。p40蛋白对应于p53 C末端的截短形式，因为它仅与识别N末端表位的单克隆抗体发生反应。这种对p53蛋白数量和长度的影响在mRNA水平上没有相关性，这表明翻译控制可能通过5'-UTR发生。我们提出了一个在这种异源环境中用于调节p53表达的UTR与部分ORF mRNA之间结构相互作用的模型。
