Relaxation helps pack DNA into a virus

Researchers at the University of California, San Diego have found that DNA packs more easily into the tight confines of a virus when given a chance to relax.

DNA is a long, unwieldy molecule that tends to repel itself because it is negatively charged, yet it can spool tightly. Within the heads of viruses, DNA can be packed to near crystalline densities, crammed in by a molecular motor.

"These are among the most powerful molecular motors we know of," says Douglas Smith, a professor of physics whose group studies them.

Within an infected cell, viruses assemble in a matter of minutes. Smith's group studies the process by isolating components of this system to watch single molecules in action.

They attach the empty head of a single virus, along with the molecular motor, to a microscopic bead that can be moved about using a laser. To another bead, they tether a molecule of viral DNA.

"It's like fishing," Smith says. "We dangle a DNA molecule in front of the viral motor. If we're lucky, the motor grabs the DNA and starts pulling it in."

Packaging proceeds in fits and starts, with slips and pauses along the way. These pauses increase, along with forces the motor counters, as the viral head becomes full.

This image shows cross-sections of the empty prohead of the bacteriophage phi29 (left) and the fully assembled virus (right). A molecular motor transports the DNA (red) into the prohead through a portal.

Scientists who model this process have had to make assumptions about the state of the DNA within. An open question is whether the DNA is in its lowest energy state, that is at equilibrium, or in a disordered configuration.

"In confinement, it could be forming all kinds of knots and tangles," said Zachary Berndsen, a graduate student in biochemistry who works with Smith. To figure this out, Berndsen stalled the motor by depriving it of chemical energy, and found that packaging rates picked up when the motor restarted. The longer the stall, the greater the acceleration.

DNA takes more than 10 minutes to fully relax inside the confines of a viral head where there's little wiggle room, the team found. That's 60,000 times as long as it takes unconfined DNA to relax.

Beyond a clearer understanding of how viruses operate, the approach offers a natural system that is a model for understanding and studying the physics of long polymers like DNA in confined spaces.

By

Keerti Mishra