Exploration of the DNA Packaging Motor of Bacteriophage phi29 for Possible Applications in Nanobiotechnology

Bacteriophage phi29 is a model organism for studies on DNA packaging, a process utilized by double-stranded DNA (dsDNA) viruses to translocate and condense their genome into the prohead. DNA packaging is accomplished by an ATP-driven motor complex that is located in a vertex of the icosahedral prohead. Biomolecular motors exhibit significant potential to be used as components in nanoelectromechanical systems and nanodevices. The DNA packaging motor of bacteriophage phi29 is particularly suitable for such explorations since it is one of the strongest biological nanomotors studied to date, it shows high processivity, it can be assembled from modified components in a defined way, it likely involves a rotary mechanism, and it can be turned on and off at will. The objective of this proposal is to gain information on how the phi29 DNA packaging motor could be used to drive synthetic nanocomponents. Specifically, this objective involves 1.) developing methods whereby synthetic particles, biomolecules, and other targets can be linked to the motor without affecting its functionality, and 2.) gaining a better understanding of the molecular working mechanism of the motor. Electron and optical microscopy and phage assembly assay of motors bearing linked targets will help determine the stoichiometry of pRNA, an essential motor component, and will conclusively determine whether the motor employs a rotary mechanism. These results will provide preliminary data informing future research on applying the phi29 DNA packaging motor in nanobiotechnology.