An Assay for Detecting RNA Topoisomerase Activity
RNA topoisomerase activity has recently been detected in multiple Type IA DNA topoisomerases from all three domains of life: bacteria, archaea, and eukarya. Many, but not all, Type IA topoisomerases are found to possess activities for not only DNA, but also RNA, suggesting that they may solve topological problems for both types of nucleic acids. Here we describe a detailed assay used by our group to detect RNA topoisomerase activity for many Type IA topoisomerases. We discuss the strategy, experimental procedures, troubleshooting, and limitations for this assay.
Key wordsTopoisomerase Top3β Circular RNA Knot
The experimental protocols described here have been previously reported in less detail in two previous publications of our group [8, 9]. This work is supported in part by the Intramural Research Program of the National Institute on Aging (Z01 AG000657-08), National Institutes of Health; the National Basic Research Program of China (2013CB911002); and National Natural Science Foundation of China (31271435).