Dependence of bacteriophage φ29 DNA packaging on ionic conditions studied by optical tweezers manipulation of single DNA molecules

Abstract

The bacteriophage φ29 portal motor is capable of packaging the φ29, 19.3 Kbp, genome to high density into its preformed capsid. The packaging process must overcome the forces due to confining the highly negative charge of the DNA to a small volume, as well as the forces due to bending the DNA on length scales smaller than one persistence length. Both of these energetic considerations can be modulated by the ionic nature of the buffer DNA packaging occurs in. To measure the effects of DNA charge shielding on the packaging process, we studied the dynamics of DNA packaging by optical tweezers in a variety of different ionic conditions. We looked at the effects monovalent, divalent, and trivalent cations have on the motor function and its dependence on external force and, we observed the rate of DNA packaging at nominal force as a function of capsid filling. Specifically, we varied the concentrations of Na+, Mg+2, and cobalt hexamine in the solution bathing the bacteriophage during packaging to see what effects, if any, these cations have. From these measurements, we present an inferred internal force as a function of percent filling of the bacteriophage capsid in a variety of ionic environments. Preliminary analysis suggests the ionic environment can modulate internal pressure, with the presence of higher valence cations better shielding the packaged DNA resulting in lower internal pressures.