Abstract
Precursor proteins used in the assembly of adenovirus virions must be processed by the virally encoded adenovirus proteinase (AVP) before the virus particle becomes infectious. An activated adenovirus proteinase, the AVP-pVIc complex, was shown to slide along viral DNA with an extremely fast one-dimensional diffusion constant, 21.0 ± 1.9 × 10(6) bp(2)/s. In principle, one-dimensional diffusion can provide a means for DNA-bound proteinases to locate and process DNA-bound substrates. Here, we show that this is correct. In vitro, AVP-pVIc complexes processed a purified virion precursor protein in a DNA-dependent reaction; in a quasi in vivo environment, heat-disrupted ts-1 virions, AVP-pVIc complexes processed five different precursor proteins in DNA-dependent reactions. Sliding of AVP-pVIc complexes along DNA illustrates a new biochemical mechanism by which a proteinase can locate its substrates, represents a new paradigm for virion maturation, and reveals a new way of exploiting the surface of DNA.
Highlights
The adenovirus proteinase and its precursor protein substrates are all sequence independent DNA binding proteins
Primary trajectory data were converted to the mean square displacement (MSD) of the distance traveled versus time, and a plot of the MSD versus time yielded a straight line whose slope is the one-dimensional diffusion constant [25, 27, 51]
Another indication the one-dimensional diffusion was due to Brownian motion was that the adenovirus proteinase (AVP)-pVIc complex displacements along DNA increased as a function of time (Fig. 2F), whereas the displacements transverse to the DNA did not (Fig. 2G), as expected for particles confined to diffuse in one dimension only, i.e. along the DNA
Summary
Mangel§4 From the ‡Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, the §Biology Department, Brookhaven National Laboratory, Upton, New York 11973, the ¶Department of Macromolecular Structures, Centro Nacional de Biotecnología (CNB-CSIC), Darwin 3, 28049 Madrid, Spain, and the ʈDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
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