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 & 9 others Chengjian Zhao, Cuisong Zhou, Binwu Ying, Bingyang Shi, Xiawei Wei, Ruotian Jiang, Lei Zhang, Guangwen Lu, JIa Geng

Research output: Contribution to journalArticleResearchpeer-review

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.
LanguageEnglish
Article number5083
Pages1-11
Number of pages11
JournalNature Communications
Volume10
DOIs
Publication statusPublished - 2019

Fingerprint

Nanopores
deoxyribonucleic acid
membranes
Membranes
DNA
delivery
Single-Stranded DNA
cells
Cell membranes
biophysics
lumens
sequencing
Cell Membrane
Biophysics
Single-Cell Analysis
cryogenics
Biological Phenomena
lipids
enzymes
insertion

Bibliographical note

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.

Cite this

Sun, K., Zhao, C., Zeng, X., Chen, Y., Jiang, X., Ding, X., ... Geng, JI. (2019). Active DNA unwinding and transport by a membrane-adapted helicase nanopore. Nature Communications, 10, 1-11. [5083]. https://doi.org/10.1038/s41467-019-13047-y
Sun, Ke ; Zhao, Changjian ; Zeng, Xiaojun ; Chen, Yuejia ; Jiang, Xin ; Ding, Xianting ; Gou, Lu ; Xie, Haiyang ; Li, Xinqiong ; Zhang, Xialin ; Lin, Sheng ; Dou, Linqin ; Wei, Long ; Niu, Haofu ; Zhang, Ming ; Tian, Ruocen ; Sawyer, Erica ; Yuan, Qingyue ; Huang, Yuqin ; Chen, Piaopiao ; Zhao, Chengjian ; Zhou, Cuisong ; Ying, Binwu ; Shi, Bingyang ; Wei, Xiawei ; Jiang, Ruotian ; Zhang, Lei ; Lu, Guangwen ; Geng, JIa. / Active DNA unwinding and transport by a membrane-adapted helicase nanopore. In: Nature Communications. 2019 ; Vol. 10. pp. 1-11.
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title = "Active DNA unwinding and transport by a membrane-adapted helicase nanopore",
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.",
author = "Ke Sun and Changjian Zhao and Xiaojun Zeng and Yuejia Chen and Xin Jiang and Xianting Ding and Lu Gou and Haiyang Xie and Xinqiong Li and Xialin Zhang and Sheng Lin and Linqin Dou and Long Wei and Haofu Niu and Ming Zhang and Ruocen Tian and Erica Sawyer and Qingyue Yuan and Yuqin Huang and Piaopiao Chen and Chengjian Zhao and Cuisong Zhou and Binwu Ying and Bingyang Shi and Xiawei Wei and Ruotian Jiang and Lei Zhang and Guangwen Lu and JIa Geng",
note = "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.",
year = "2019",
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Sun, K, Zhao, C, Zeng, X, Chen, Y, Jiang, X, Ding, X, Gou, L, Xie, H, Li, X, Zhang, X, Lin, S, Dou, L, Wei, L, Niu, H, Zhang, M, Tian, R, Sawyer, E, Yuan, Q, Huang, Y, Chen, P, Zhao, C, Zhou, C, Ying, B, Shi, B, Wei, X, Jiang, R, Zhang, L, Lu, G & Geng, JI 2019, 'Active DNA unwinding and transport by a membrane-adapted helicase nanopore', Nature Communications, vol. 10, 5083, pp. 1-11. https://doi.org/10.1038/s41467-019-13047-y

Active DNA unwinding and transport by a membrane-adapted helicase nanopore. / Sun, Ke; Zhao, Changjian; Zeng, Xiaojun; Chen, Yuejia; Jiang, Xin ; Ding, Xianting; Gou, Lu; Xie, Haiyang ; Li, Xinqiong; Zhang, Xialin; Lin, Sheng; Dou, Linqin; Wei, Long; Niu, Haofu; Zhang, Ming; Tian, Ruocen; Sawyer, Erica; Yuan, Qingyue; Huang, Yuqin; Chen, Piaopiao; Zhao, Chengjian; Zhou, Cuisong; Ying, Binwu; Shi, Bingyang; Wei, Xiawei; Jiang, Ruotian; Zhang, Lei; Lu, Guangwen; Geng, JIa.

In: Nature Communications, Vol. 10, 5083, 2019, p. 1-11.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Sun, Ke

AU - Zhao, Changjian

AU - Zeng, Xiaojun

AU - Chen, Yuejia

AU - Jiang, Xin

AU - Ding, Xianting

AU - Gou, Lu

AU - Xie, Haiyang

AU - Li, Xinqiong

AU - Zhang, Xialin

AU - Lin, Sheng

AU - Dou, Linqin

AU - Wei, Long

AU - Niu, Haofu

AU - Zhang, Ming

AU - Tian, Ruocen

AU - Sawyer, Erica

AU - Yuan, Qingyue

AU - Huang, Yuqin

AU - Chen, Piaopiao

AU - Zhao, Chengjian

AU - Zhou, Cuisong

AU - Ying, Binwu

AU - Shi, Bingyang

AU - Wei, Xiawei

AU - Jiang, Ruotian

AU - Zhang, Lei

AU - Lu, Guangwen

AU - Geng, JIa

N1 - 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.

PY - 2019

Y1 - 2019

N2 - 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.

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