Abstract
SIMIAN virus 40 (SV40) replicates in nuclei of human and monkey cells. One viral protein, large tumour (T) antigen, is required for the initiation of DNA replication1,2. The development of in vitro replication systems which retain this property has facilitated the identification of the cellular components required for replication3–5. T antigen recognizes the pentanucleotide 5′-GAGGC-3′ which is present in four copies6,7 within the 64 base-pairs (bp) of the core origin8,9. In the presence of ATP it binds with increased affinity10,11 forming a distinctive, bilobed structure visible in electron micrographs12. As a helicase13, it unwinds SV40 DNA bidirectional ly from the origin14–16. We report here that in vitro and in the presence of ATP, T antigen assembles a double hexamer, centred on the core origin and extending beyond it by 12 bp in each direction. The assembly of this dodecamer initiates an untwisting of the duplex by 2–3 turns. In the absence of ATP, a tetrameric structure is the largest found at the core origin. In the absence of DNA, but in the presence of ATP or its non-hydrolysable analogues, T antigen assembles into hexamers. This suggests that ATP effects an allosteric change in the monomer. The change alters protein-protein interactions and allows the assembly of a double hexamer, which initiates replication at the core origin.
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Mastrangelo, I., Hough, P., Wall, J. et al. ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication. Nature 338, 658–662 (1989). https://doi.org/10.1038/338658a0
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DOI: https://doi.org/10.1038/338658a0
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