Graduate Thesis Or Dissertation
 

Adenovirus replication in trans : a new replication pathway

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/d504rn70q

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  • Replication of plasmid molecules containing either a left-end or a right-end adenovirus origin was investigated. When the plasmids were linearized to expose their respective adenovirus termini, mixed, and transfected with helper adenovirus DNA into the human 293 cell line, a new molecule was detected. The new molecule was the size predicted for a recombinant between the two input plasmids. The recombinational event, however, was not due to simple homologous recombination and was totally dependent upon adenovirus replication. No molecules of the same size were detected in the absence of helper adenovirus DNA, indicating that detection of the new molecule was enabled by a function provided by the adenovirus DNA, most likely replication. The molecules appeared to be replicating by an adenovirus-driven mechanism as indicated by their covalent attachment to protein and their accumulation from 35-72 hours in a time course experiment. The available evidence indicates that the molecules were produced by replication followed by recombination rather than vice-versa. A mechanism, called trans replication, is postulated whereby the displaced complementary strands produced during replication interact via base pairing to form a duplex. Kinetic data of adenovirus replication previously obtained by other workers is consistent with the process functioning in the normal adenovirus replication cycle. Such a process is a hitherto unknown means for adenovirus to finish complementary replication. As such, it represents the first clear violation of the Meselson-Stahl principle of semi-conservative replication that has been observed in any biological system.
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