Kotkowiak Weronika, Pasternak Anna, Kierzek Ryszard
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland.
PLoS One. 2016 Feb 1;11(2):e0148282. doi: 10.1371/journal.pone.0148282. eCollection 2016.
In this study, several RNA polymerases were used for the first time to examine the possibility of transcriptional incorporation of 5'-N-triphosphates of 5'-amino-5'-deoxyribonucleosides (5'NH NTPs). The T3, T7, Sp6 and T7 Y639F RNA polymerases were employed to show that the full-length transcript cannot be synthesized. The results suggest that the application of 5'NH NTPs could decrease transcription reaction rates. What is more, the modification of transcription conditions had no influence on the rate of 5'NH NTPs incorporation. Based on experimental data it is postulated that 5'NH NTPs can be used as potential transcription inhibitors. Our findings expand the knowledge on suitable uses of the 5'-N-triphosphates of 5'-amino-5'-deoxyribonucleoside and the exact mechanism of transcriptional inhibition.
在本研究中,首次使用了几种RNA聚合酶来检测5'-氨基-5'-脱氧核糖核苷的5'-N-三磷酸(5'NH NTPs)转录掺入的可能性。使用T3、T7、Sp6和T7 Y639F RNA聚合酶来表明无法合成全长转录本。结果表明,5'NH NTPs的应用会降低转录反应速率。此外,转录条件的改变对5'NH NTPs的掺入速率没有影响。基于实验数据推测,5'NH NTPs可作为潜在的转录抑制剂。我们的研究结果扩展了关于5'-氨基-5'-脱氧核糖核苷的5'-N-三磷酸的合适用途以及转录抑制的确切机制的知识。