Issue 10, 2010

Robust properties of membrane-embedded connector channel of bacterial virus phi29 DNA packagingmotor

Abstract

Biological systems contain highly-ordered macromolecular structures with diverse functions, inspiring their utilization in nanotechnology. A motor allows linear dsDNA viruses to package their genome into a preformed procapsid. The central component of the motor is the portal connector that acts as a pathway for the translocation of dsDNA. The elegant design of the connector and its channel motivates its application as an artificial nanopore (Nature Nanotechnology, 4, 765–772). Herein, we demonstrate the robust characteristics of the connector of the bacteriophage phi29 DNA packaging motor by single pore electrophysiological assays. The conductance of each pore is almost identical and is perfectly linear with respect to the applied voltage. Numerous transient current blockade events induced by dsDNA are consistent with the dimensions of the channel and dsDNA. Furthermore, the connector channel is stable under a wide range of experimental conditions including high salt and pH 2–12. The robust properties of the connector nanopore made it possible to develop a simple reproducible approach for connector quantification. The precise number of connectors in each sheet of the membrane was simply derived from the slopes of the plot of voltage against current. Such quantifications led to a reliable real time counting of DNA passing through the channel. The fingerprint of DNA translocation in this system has provided a new tool for future biophysical and physicochemical characterizations of DNA transportation, motion, and packaging.

Graphical abstract: Robust properties of membrane-embedded connector channel of bacterial virus phi29 DNA packaging motor

Supplementary files

Article information

Article type
Paper
Submitted
12 Feb 2010
Accepted
07 Apr 2010
First published
04 Jun 2010

Mol. BioSyst., 2010,6, 1844-1852

Robust properties of membrane-embedded connector channel of bacterial virus phi29 DNA packaging motor

P. Jing, F. Haque, A. P. Vonderheide, C. Montemagno and P. Guo, Mol. BioSyst., 2010, 6, 1844 DOI: 10.1039/C003010D

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Spotlight

Advertisements