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Vol. 273, Issue 2, 1165-1174, January 9, 1998

Initiation of DNA Replication at Palindromic Telomeres Is Mediated by a Duplex-to-Hairpin Transition Induced by the Minute Virus of Mice Nonstructural Protein NS1

From the Department of Applied Tumor Virology, Abteilung 0610, Deutsches Krebsforschungszentrum and Formation INSERM U375, D-69009 Heidelberg, Germany

The linear single-stranded DNA genome of the minute virus of mice (MVM) is replicated via a double-stranded replicative form (RF) intermediate. Amplification of this RF is initiated by the folding-back of palindromic sequences serving as primers for strand-displacement synthesis and formation of dimeric RF DNA. Using an in vitro replication assay and a cloned MVM DNA template, we observed hairpin-primed DNA replication at both MVM DNA termini, with a bias toward right-end initiation. Initiation of DNA replication is favored by nuclear components of A9 cell extract and highly stimulated by the MVM nonstructural protein NS1. Hairpin-primed DNA replication is also observed in the presence of NS1 and the Klenow fragment of the Escherichia coli DNA polymerase I. Addition of ATPS (adenosine 5-O-(thiotriphosphate)) blocks the initiation of DNA replication but not the extension of pre-existing hairpin primers formed in the presence of NS1 only. The NS1-mediated unwinding of the right-end palindrome may account for the recently reported capacity of NS1 for driving dimer RF synthesis in vitro.

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