Abstract
Electron microscope autoradiograms revealed that grains (viral DNA) were found in the nuclei of cells coinfected with adeno-associated virus type 1 (AAV1) and a temperature-sensitive mutant of human adenovirus type 31 (H31 tsA13) at the nonpermissive temperature (40°). Nuclei isolated from cells coinfected with AAV1 and H31 tsA13 at 40° were active in AAV1-DNA synthesis in vitro. Alkaline sucrose gradient analysis of the in vitro products showed that DNA synthesized with nuclei isolated from coinfected cells sedimented faster than marker AAV1-DNA, although DNA synthesized with nuclei isolated from cells coinfected in the presence of cycloheximide cosedimented with the marker DNA. This fast-sedimenting DNA shifted to the position of the marker DNA after treatment with papain, trypsin, and sodium dodecyl sulfate followed by extraction with phenol. This observation suggests the involvement of a protein in the formation of fast-sedimenting viral DNA.
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