Graduate Thesis Or Dissertation
 

Nucleoprotein structure of adenovirus

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  • The structure of the nucleoprotein core of adenovirus has been investigated. Using a combination of biophysical techniques and nuclease digestion I have determined that the adenovirus core, as isolated with pyridine, shares several physical properties with cellular chromatin. The most notable difference is the lack of a nucleosome repeat in the staph nuclease digestion intermediates from adenovirus cores. I have exploited this difference to study the fate of the nucleoprotein structure of adenovirus DNA early during productive infection. When nuclei from cells infected with ³²P-labeled adenovirus are digested with staph nuclease the pattern of labeled digestion intermediates more closely resembles the pattern of cellular intermediates. This result indicates that virus core proteins are replaced, early during infection, with cellular histones. The newly assembled adenovirus nucleosomes differ from the bulk of the cellular chromatin by having a shorter repeat (160 vs. 185 base pairs) and more labile linker and core regions. Experiments with inhibitors of cellular DNA synthesis indicate that adenovirus nucleosomes are assembled independently of cellular. DNA synthesis. Using the Southern blotting technique it is shown that newly synthesized adenovirus DNA is not in a nucleosome repeat late in infection. At this time of infection, however, the input adenovirus genomes persist in a nucleosome structure. The results of these experiments are discussed in terms of the template requirements for eucaryotic transcription and replication.
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