Mechanochemistry of a Viral DNA Packaging Motor |
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Authors: | Jin Yu Craig L. Hetherington George Oster |
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Affiliation: | 1 Department of Physics, University of California, Berkeley, CA 94720, USA 2 Department of Physics, Chemistry, Molecular and Cell Biology, Howard Hughes Medical Insititute, University of California, Berkeley, CA 94720, USA 3 Department of Molecular and Cell Biology, Environmental Science, Policy, & Management, University of California, Berkeley, CA 94720, USA |
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Abstract: | The pentameric ATPase motor gp16 packages double-stranded DNA into the bacteriophage ?29 virus capsid. On the basis of the results of single-molecule experimental studies, we propose a push and roll mechanism to explain how the packaging motor translocates the DNA in bursts of four 2.5 bp power strokes, while rotating the DNA. In this mechanism, each power stroke accompanies Pi release after ATP hydrolysis. Since the high-resolution structure of the gp16 motor is not available, we borrowed characterized features from the P4 RNA packaging motor in bacteriophage ?12. For each power stroke, a lumenal lever from a single subunit is electrostatically steered to the DNA backbone. The lever then pushes sterically, orthogonal to the backbone axis, such that the right-handed DNA helix is translocated and rotated in a left-handed direction. The electrostatic association allows tight coupling between the lever and the DNA and prevents DNA from slipping back. The lever affinity for DNA decreases towards the end of the power stroke and the DNA rolls to the lever on the next subunit. Each power stroke facilitates ATP hydrolysis in the next catalytic site by inserting an Arg -finger into the site, as captured in ?12-P4. At the end of every four power strokes, ADP release happens slowly, so the cycle pauses constituting a dwell phase during which four ATPs are loaded into the catalytic sites. The next burst phase of four power strokes starts once spontaneous ATP hydrolysis takes place in the fifth site without insertion of an Arg finger. The push and roll model provides a new perspective on how a multimeric ATPase transports DNA, and it might apply to other ring motors as well. |
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Keywords: | dsDNA, double-stranded DNA cryo-EM, cryo-electron microscope |
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