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 Chen, Chengjian Zhao, Cuisong Zhou, Binwu Ying, Bingyang Shi, Xiawei Wei, Ruotian Jiang, Lei Zhang (), Guangwen Lu () and Jia Geng ()
Additional contact information
Ke Sun: Sichuan University and Collaborative Innovation Center for Biotherapy
Changjian Zhao: Sichuan University and Collaborative Innovation Center for Biotherapy
Xiaojun Zeng: Sichuan University and Collaborative Innovation Center for Biotherapy
Yuejia Chen: Sichuan University and Collaborative Innovation Center for Biotherapy
Xin Jiang: Sichuan University and Collaborative Innovation Center for Biotherapy
Xianting Ding: Shanghai Jiao Tong University
Lu Gou: Xi’an Jiaotong University
Haiyang Xie: Shanghai Jiao Tong University
Xinqiong Li: Sichuan University and Collaborative Innovation Center for Biotherapy
Xialin Zhang: Sichuan University and Collaborative Innovation Center for Biotherapy
Sheng Lin: Sichuan University and Collaborative Innovation Center for Biotherapy
Linqin Dou: Sichuan University and Collaborative Innovation Center for Biotherapy
Long Wei: Sichuan University and Collaborative Innovation Center for Biotherapy
Haofu Niu: Sichuan University and Collaborative Innovation Center for Biotherapy
Ming Zhang: Sichuan University and Collaborative Innovation Center for Biotherapy
Ruocen Tian: Sichuan University and Collaborative Innovation Center for Biotherapy
Erica Sawyer: Sichuan University and Collaborative Innovation Center for Biotherapy
Qingyue Yuan: Sichuan University and Collaborative Innovation Center for Biotherapy
Yuqin Huang: Sichuan University
Piaopiao Chen: Sichuan University and Collaborative Innovation Center for Biotherapy
Chengjian Zhao: Sichuan University and Collaborative Innovation Center for Biotherapy
Cuisong Zhou: Sichuan University
Binwu Ying: Sichuan University and Collaborative Innovation Center for Biotherapy
Bingyang Shi: Henan University
Xiawei Wei: Sichuan University and Collaborative Innovation Center for Biotherapy
Ruotian Jiang: Sichuan University
Lei Zhang: Xi’an Jiaotong University
Guangwen Lu: Sichuan University
Jia Geng: Sichuan University and Collaborative Innovation Center for Biotherapy

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: Abstract 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.

Date: 2019
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DOI: 10.1038/s41467-019-13047-y

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