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

Author

Listed:
  • 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)

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.

Suggested Citation

  • 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 & Ruoc, 2019. "Active DNA unwinding and transport by a membrane-adapted helicase nanopore," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-13047-y
    DOI: 10.1038/s41467-019-13047-y
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    Cited by:

    1. Xiaojun Wei & Xiaoqin Wang & Zehui Zhang & Yuanyuan Luo & Zixin Wang & Wen Xiong & Piyush K. Jain & John R. Monnier & Hui Wang & Tony Y. Hu & Chuanbing Tang & Helmut Albrecht & Chang Liu, 2022. "A click chemistry amplified nanopore assay for ultrasensitive quantification of HIV-1 p24 antigen in clinical samples," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    2. Shiyan Liu & Mutian Chen & Yichang Wang & Yuqing Lei & Ting Huang & Yabin Zhang & Sin Man Lam & Huihui Li & Shiqian Qi & Jia Geng & Kefeng Lu, 2023. "The ER calcium channel Csg2 integrates sphingolipid metabolism with autophagy," Nature Communications, Nature, vol. 14(1), pages 1-19, December.

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