title: Structure and inhibition of herpesvirus DNA packaging terminase nuclease domain.
abstract: During viral replication, herpesviruses package their DNA into the procapsid by means of the terminase protein complex. In human cytomegalovirus (herpesvirus 5), the terminase is composed of subunits UL89 and UL56. UL89 cleaves the long DNA concatemers into unit-length genomes of appropriate length for encapsidation. We used ESPRIT, a high-throughput screening method, to identify a soluble purifiable fragment of UL89 from a library of 18,432 randomly truncated ul89 DNA constructs. The purified protein was crystallized and its three-dimensional structure was solved. This protein corresponds to the key nuclease domain of the terminase and shows an RNase H/integrase-like fold. We demonstrate that UL89-C has the capacity to process the DNA and that this function is dependent on Mn(2+) ions, two of which are located at the active site pocket. We also show that the nuclease function can be inactivated by raltegravir, a recently approved anti-AIDS drug that targets the HIV integrase.
authors.affiliation: Institute for Research in Biomedicine, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Nadal
authors.forename: Philippe J
authors.initials: PJ
authors.lastname: Mas
authors.forename: Phillipe J
authors.initials: PJ
authors.lastname: Mas
authors.forename: Alexandre G
authors.initials: AG
authors.lastname: Blanco
authors.forename: Carme
authors.initials: C
authors.lastname: Arnan
authors.forename: Maria
authors.initials: M
authors.lastname: Solà
authors.forename: Darren J
authors.initials: DJ
authors.lastname: Hart
authors.forename: Miquel
authors.initials: M
authors.lastname: Coll
mesh_terms: D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003587:Cytomegalovirus / Q000201:enzymology* / Q000235:genetics; D042003:DNA Packaging; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D004279:DNA, Viral; C436448:UL89 protein, Cytomegalovirus; D014764:Viral Proteins
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