Active DNA unwinding and transport by a membrane-adapted helicase nanopore

Ke Sun, Changjian Zhao, Xiaojun Zeng, Yuejia Chen, Xin Jiang, Xianting Ding, Lu Gou, Haiyang Xie, Xinqiong Li, Xialin Zhang, Sheng Lin, Linqin Dou, Long Wei, Haofu Niu, Ming Zhang, Ruocen Tian, Erica Sawyer, Qingyue Yuan, Yuqin Huang, Piaopiao ChenChengjian Zhao, Cuisong Zhou, Binwu Ying, Bingyang Shi, Xiawei Wei, Ruotian Jiang, Lei Zhang, Guangwen Lu, JIa Geng

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Nanoscale transport through nanopores and live-cell membranes plays a vital role in both key biological processes as well as biosensing and DNA sequencing. Active translocation of DNA through these nanopores usually needs enzyme assistance. Here we present a nanopore derived from truncated helicase E1 of bovine papillomavirus (BPV) with a lumen diameter of c.a. 1.3 nm. Cryogenic electron microscopy (cryo-EM) imaging and single channel recording confirm its insertion into planar lipid bilayer (BLM). The helicase nanopore in BLM allows the passive single-stranded DNA (ssDNA) transport and retains the helicase activity in vitro. Furthermore, we incorporate this helicase nanopore into the live cell membrane of HEK293T cells, and monitor the ssDNA delivery into the cell real-time at single molecule level. This type of nanopore is expected to provide an interesting tool to study the biophysics of biomotors in vitro, with potential applications in biosensing, drug delivery and real-time single cell analysis.
Original languageEnglish
Article number5083
Pages (from-to)1-11
Number of pages11
JournalNature Communications
Publication statusPublished - 2019

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Copyright the Author(s) 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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