Guerrero-González Maria L, Rodríguez-Kessler Margarita, Jiménez-Bremont Juan F
Division de Biologia Molecular, Instituto Potosino de Investigacion Cientifica y Tecnologica, Camino a la Presa de San Jose 2055, AP 3-74 Tangamanga, 78216, San Luis Potosi, SLP, Mexico.
Mol Biol Rep. 2014;41(4):2427-43. doi: 10.1007/s11033-014-3098-5. Epub 2014 Jan 17.
The translational efficiency of an mRNA can be modulated by elements located in the 5'-untranslated region. The flavin-containing polyamine oxidases catabolize oxidative deamination of spermidine and spermine, thus contributing to polyamine homeostasis as well as diverse biological processes through their reaction products. In this study, we characterized the uORF of AtPAO2 gene using the GUS reporter gene. Transgenic lines harboring the native AtPAO2 promoter or the constitutive CaMV 35S promoter show that the uORF negatively affects GUS expression. Exogenous applications of PAs positively modulate GUS expression, thus alleviating the negative effect of AtPAO2 uORF, while treatments with MGBG inhibitor show an opposite effect. Our data suggest that AtPAO2 uORF regulatory mechanism is modulated by polyamines. In addition, we present a comparative in silico study of the uORFs identified in several plant transcripts encoding polyamine oxidases in both mono- and dicotyledonous plants as well as in the Bryophyte Physcomitrella patens. The polyamine oxidase uORF-encoded peptides are conserved among families and share conserved features such as their position, length, and amino acid sequence. Our findings provide new insights into the regulatory mechanism of polyamine oxidase genes and encourage further exploration to assess the biological significance of uORFs in the polyamine catabolic pathway.
mRNA的翻译效率可由位于5'-非翻译区的元件调节。含黄素的多胺氧化酶催化亚精胺和精胺的氧化脱氨反应,通过其反应产物维持多胺稳态并参与多种生物学过程。在本研究中,我们利用GUS报告基因对AtPAO2基因的上游开放阅读框进行了表征。携带天然AtPAO2启动子或组成型CaMV 35S启动子的转基因株系表明,该上游开放阅读框对GUS表达有负面影响。外源施加多胺可正向调节GUS表达,从而减轻AtPAO2上游开放阅读框的负面影响,而用MGBG抑制剂处理则显示出相反的效果。我们的数据表明,AtPAO2上游开放阅读框的调控机制受多胺调节。此外,我们对单子叶植物、双子叶植物以及苔藓植物小立碗藓中几种编码多胺氧化酶的植物转录本中鉴定出的上游开放阅读框进行了比较计算机模拟研究。多胺氧化酶上游开放阅读框编码的肽在不同家族间具有保守性,并具有如位置、长度和氨基酸序列等保守特征。我们的研究结果为多胺氧化酶基因的调控机制提供了新的见解,并鼓励进一步探索以评估上游开放阅读框在多胺分解代谢途径中的生物学意义。