Vaccinia virus (VACV), the prototype member of the Poxviridae, replicates in the cytoplasm of an infected cell. The catalytic subunit of the DNA polymerase E9 binds the heterodimeric processivity factor A20/D4 to form the functional polymerase holoenzyme. Here we present the crystal structure of full-length E9 at 2.7 Å resolution that permits identification of important poxvirus-specific structural insertions. One insertion in the palm domain interacts with C-terminal residues of A20 and thus serves as the processivity factor-binding site. This is in strong contrast to all other family B polymerases that bind their co-factors at the C terminus of the thumb domain. The VACV E9 structure also permits rationalization of polymerase inhibitor resistance mutations when compared with the closely related eukaryotic polymerase delta-DNA complex.

BioCeV Institute of Microbiology Czech Academy of Sciences Prumyslova 595 252 50 Vestec Czech Republic

Emerging Pathogens Laboratory Fondation Mérieux 21 Avenue Tony Garnier 69007 Lyon France

Faculty of Science Charles University Hlavova 8 128 43 Prague 2 Czech Republic

Institut de Biologie Structurale Université Grenoble Alpes CNRS CEA 71 Avenue des Martyrs 38042 Grenoble France

Integrated Structural Biology Grenoble CNRS CEA Université Grenoble Alpes EMBL 71 Avenue des Martyrs 38042 Grenoble France

Unité de Virologie Institut de Recherche Biomédicale des Armées BP 73 91223 Brétigny sur Orge Cedex France

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Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance

Meeting report: 32nd International Conference on Antiviral Research