Engineering of the Fluorescent-energy-conversion Arm of Phi29 DNA Packaging Motor for Single-molecule Studies

Overview

Journal Small

Date 2009 Sep 11

PMID 19743427

Citations 11

Authors

Tae Jin Lee

Hui Zhang

Chun-Li Chang

Cagri Savran

Peixuan Guo

Affiliations

Soon will be listed here.

Abstract

The bacteriophage phi29 DNA packaging motor contains a protein core with a central channel comprising twelve copies of re-engineered gp10 protein geared by six copies of packaging RNA (pRNA) and a DNA packaging protein gp16 with unknown copies. Incorporation of this nanomotor into a nanodevice would be beneficial for many applications. To this end, extension and modification of the motor components are necessary for the linkage of this motor to other nanomachines. Here the re-engineering of the motor DNA packaging protein gp16 by extending its length and doubling its size using a fusion protein technique is reported. The modified motor integrated with the eGFP-gp16 maintains the ability to convert the chemical energy from adenosine triphosphate (ATP) hydrolysis to mechanical motion and package DNA. The resulting DNA-filled capsid is subsequently converted into an infectious virion. The extended part of the gp16 arm is a fluorescent protein eGFP, which serves as a marker for tracking the motor in single-molecule studies. The activity of the re-engineered motor with eGFP-gp16 is also observed directly with a bright-field microscope via its ability to transport a 2-microm-sized cargo bound to the DNA.

Citing Articles

Revolving ATPase motors as asymmetrical hexamers in translocating lengthy dsDNA via conformational changes and electrostatic interactions in phi29, T7, herpesvirus, mimivirus, , and .

Weitao T, Grandinetti G, Guo P Exploration (Beijing). 2023; 3(2):20210056.

PMID: 37324034 PMC: 10191066. DOI: 10.1002/EXP.20210056.

Optical tweezer and TIRF microscopy for single molecule manipulation of RNA/DNA nanostructures including their rubbery property and single molecule counting.

Ghimire C, Guo P Biophys Rep. 2023; 7(6):449-474.

PMID: 37288367 PMC: 10210058. DOI: 10.52601/bpr.2021.210003.

Advanced Nanoscale Approaches to Single-(Bio)entity Sensing and Imaging.

Neves M, Martin-Yerga D Biosensors (Basel). 2018; 8(4).

PMID: 30373209 PMC: 6316691. DOI: 10.3390/bios8040100.

Construction of Asymmetrical Hexameric Biomimetic Motors with Continuous Single-Directional Motion by Sequential Coordination.

Zhao Z, Zhang H, Shu D, Montemagno C, Ding B, Li J Small. 2016; 13(1).

PMID: 27709780 PMC: 5217803. DOI: 10.1002/smll.201601600.

An Arginine Finger Regulates the Sequential Action of Asymmetrical Hexameric ATPase in the Double-Stranded DNA Translocation Motor.

Zhao Z, De-Donatis G, Schwartz C, Fang H, Li J, Guo P Mol Cell Biol. 2016; 36(19):2514-23.

PMID: 27457616 PMC: 5021374. DOI: 10.1128/MCB.00142-16.

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