An Assay for Detecting RNA Topoisomerase Activity

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1703)

Abstract

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 words

Topoisomerase Top3β Circular RNA Knot 

Notes

Acknowledgment

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).

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Genome Instability and Chromatin Remodeling Section, Lab of Genetics and GenomicsNational Institute on Aging, National Institute on HealthBaltimoreUSA
  2. 2.State Key Laboratory of Protein and Plant Gene ResearchSchool of Life Sciences, Peking UniversityBeijingChina

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