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.

Abstract

Active translocation of DNA through nanopores usually needs enzyme assistance. Here authors present a nanopore derived from helicase E1 of bovine papillomavirus (BPV) which acts as a conductive pore embedded in lipid membrane to allow the translocation of ssDNA and unwinding of dsDNA.

Contributors

Lei Zhang:

ORCID: http://orcid.org/0000-0002-4880-824X

[e-mail]

Guangwen Lu:

ORCID: http://orcid.org/0000-0001-7568-592X

[e-mail]

Jia Geng: [e-mail]

Journal

Journal ID (nlm-ta): Nat Commun

Journal ID (iso-abbrev): Nat Commun

Title: Nature Communications

Publisher: Nature Publishing Group UK (London )

ISSN (Electronic): 2041-1723

Publication date (Electronic): 8 November 2019

Publication date PMC-release: 8 November 2019

Publication date Collection: 2019

Volume: 10

Affiliations

[1 ]ISNI 0000 0001 0807 1581, GRID grid.13291.38, Department of Laboratory Medicine, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, , Sichuan University and Collaborative Innovation Center for Biotherapy, ; Chengdu, 610041 China

[2 ]ISNI 0000 0004 0368 8293, GRID grid.16821.3c, Institute for Personalized Medicine, State Key Laboratory of Oncogenes and Related Genes, School of Biomedical Engineering, , Shanghai Jiao Tong University, ; Shanghai, 200030 China

[3 ]ISNI 0000 0001 0599 1243, GRID grid.43169.39, MOE Key Laboratory for Nonequilibrium Synthesis and Modulation of Condensed Matter, School of Science, , Xi’an Jiaotong University, ; Xi’an, 710049 China

[4 ]ISNI 0000 0001 0807 1581, GRID grid.13291.38, College of Chemistry, , Sichuan University, ; Chengdu, 610041 China

[5 ]ISNI 0000 0000 9139 560X, GRID grid.256922.8, Henan-Macquarie Uni Joint Center for Biomedical Innovation, School of Life Sciences, , Henan University, ; Kaifeng, Henan 475004 China

[6 ]ISNI 0000 0001 0807 1581, GRID grid.13291.38, Laboratory of Anesthesia and Critical Care Medicine, Department of Anesthesiology, West China Hospital, , Sichuan University, ; Chengdu, Sichuan 610000 China

[7 ]ISNI 0000 0001 0807 1581, GRID grid.13291.38, West China Hospital Emergency Department (WCHED), State Key Laboratory of Biotherapy, West China Hospital, , Sichuan University, ; Chengdu, Sichuan 610041 China

Article

Publisher ID: 13047

DOI: 10.1038/s41467-019-13047-y

PMC ID: 6841704

PubMed ID: 31704937

Copyright statement: © The Author(s) 2019

License:

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Funding

Funded by: FundRef https://doi.org/10.13039/501100001809, National Natural Science Foundation of China (National Science Foundation of China);

Award ID: 81541133

Award Recipient : Jia Geng

Categories

Subject: Article

Custom metadata

issue-copyright-statement © The Author(s) 2019