Abstract
Initiation of Adenovirus DNA replicationin vitro requires the presence of three viral proteins (pTP, pol, DBP) and two cellular transcription factors, NFI and Oct-1, that stimulate replication more than 100-fold. NFI assists in binding and positioning of the DNA polymerase in the origin whereas Oct-1 changes the structure of origin DNA. Optimal templates contain, in addition to origin sequences, the covalently bound viral terminal protein (TP). This terminal protein stimulates the template activity over 20 fold compared to protein-free templates. To study the way in which TP exerts its functionin vitro we devised a novel method to isolate and label a short origin containing fragment in which the TP was bound in a functional form. This fragment replicated very efficiently and could be used for studying the binding of other replication proteins. Employing α-chymotrypsin digestion we show that for enhancement of replicationin vitro only a small part of TP is required.
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Abbreviations: Adenovirus, DNA replication, terminal protein, transcription factors.
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Pronk, R., Stuiver, M.H. & van der Vliet, P.C. Adenovirus DNA replication: The function of the covalently bound terminal protein. Chromosoma 102 (Suppl 1), S39–S45 (1992). https://doi.org/10.1007/BF02451784
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DOI: https://doi.org/10.1007/BF02451784