A step towards in vitro reconstruction of the replication of Herpes Simplex Virus-1 (2014)

Undergraduates: Molly Laux, Dr. Oya Bermek, Dr. Jack D Griffith, Smaranda Willcox, Sezgin ¿zg¿r

Faculty Advisor: Jack Griffith
Department: Chemistry

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Herpes Simplex Virus-1 (HSV-1) is a valuable model for eukaryotic DNA replication as the replication machinery consists of only seven viral proteins and they can be all purified from insect cells. This includes the origin-binding protein UL9, which acts as a DNA helicase as well as a DNA-dependent NTPase. The UL9 binding ability to DNA, both alone and in protein complexes, was visualized through electron microscopy. In this study, we showed that UL9 binds to 74% of the DNA molecules containing the minimal HSV-1 origin of replication, in presence of single-strand binding protein, ICP8. Furthermore, addition of ATP leads to the changes in the structure of the supertwisted DNA. The optimization of the binding conditions at the replication origin, prior to the addition of the complete set of replication proteins will provide the first step in the reconstruction of HSV-1 replication.