Ionic effects on viral DNA packaging and portal motor function in bacteriophage phi 29.
In many viruses, DNA is confined at such high density that its bending rigidity and electrostatic self-repulsion present a strong energy barrier in viral assembly. Therefore, a powerful molecular motor is needed to package the DNA into the viral capsid. Here, we investigate the role of electrostatic repulsion on single DNA packaging dynamics in bacteriophage phi 29 via optical tweezers measurements. We show that ionic screening strongly affects the packing forces, confirming the importance of electrostatic repulsion. Separately, we find that ions affect the motor function. We separate these effects through constant force measurements and velocity versus load measurements at both low and high capsid filling. Regarding motor function, we find that eliminating free Mg(2+) blocks initiation of packaging. In contrast, Na(+) is not required, but it increases the motor velocity by up to 50% at low load. Regarding internal resistance, we find that the internal force was lowest when Mg(2+) was the dominant ion or with the addition of 1 mM Co(3+). Forces resisting DNA confinement were up to approximately 80% higher with Na(+) as the dominant counterion, and only approximately 90% of the genome length could be packaged in this condition. The observed trend of the packing forces is in accord with that predicted by DNA charge-screening theory. However, the forces are up to six times higher than predicted by models that assume coaxial spooling of the DNA and interaction potentials derived from DNA condensation experiments. The forces are also severalfold higher than ejection forces measured with bacteriophage lambda.
Fuller
Derek N
DN
Department of Physics, University of California, San Diego, Mail Code 0379, 9500 Gilman Drive, La Jolla, CA 92093, USA.
Rickgauer
John Peter
JP
Jardine
Paul J
PJ
Grimes
Shelley
S
Anderson
Dwight L
DL
Smith
Douglas E
DE
D017103:Bacillus Phages / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D002213:Capsid / Q000737:chemistry / Q000502:physiology; D002413:Cations, Divalent / Q000737:chemistry; D002414:Cations, Monovalent / Q000737:chemistry; D003035:Cobalt / Q000737:chemistry / Q000502:physiology; D004279:DNA, Viral / Q000737:chemistry / Q000502:physiology*; D008274:Magnesium / Q000737:chemistry / Q000502:physiology; D052898:Optical Tweezers; D011237:Predictive Value of Tests; D012964:Sodium / Q000737:chemistry / Q000502:physiology; D055672:Static Electricity; D019065:Virus Assembly / Q000502:physiology*
D002413:Cations, Divalent; D002414:Cations, Monovalent; D004279:DNA, Viral; D003035:Cobalt; D012964:Sodium; D008274:Magnesium