title: Crystal structure of the gamma-2 herpesvirus LANA DNA binding domain identifies charged surface residues which impact viral latency.
abstract: Latency-associated nuclear antigen (LANA) mediates γ2-herpesvirus genome persistence and regulates transcription. We describe the crystal structure of the murine gammaherpesvirus-68 LANA C-terminal domain at 2.2 Å resolution. The structure reveals an alpha-beta fold that assembles as a dimer, reminiscent of Epstein-Barr virus EBNA1. A predicted DNA binding surface is present and opposite this interface is a positive electrostatic patch. Targeted DNA recognition substitutions eliminated DNA binding, while certain charged patch mutations reduced bromodomain protein, BRD4, binding. Virus containing LANA abolished for DNA binding was incapable of viable latent infection in mice. Virus with mutations at the charged patch periphery exhibited substantial deficiency in expansion of latent infection, while central region substitutions had little effect. This deficiency was independent of BRD4. These results elucidate the LANA DNA binding domain structure and reveal a unique charged region that exerts a critical role in viral latent infection, likely acting through a host cell protein(s).
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Correia
authors.forename: Sofia A
authors.initials: SA
authors.lastname: Cerqueira
authors.forename: Chantal
authors.initials: C
authors.lastname: Beauchemin
authors.forename: Marta
authors.initials: M
authors.lastname: Pires de Miranda
authors.forename: Shijun
authors.initials: S
authors.lastname: Li
authors.forename: Rajesh
authors.initials: R
authors.lastname: Ponnusamy
authors.forename: Lénia
authors.initials: L
authors.lastname: Rodrigues
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Schneider
authors.forename: Maria A
authors.initials: MA
authors.lastname: Carrondo
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Kaye
authors.forename: J Pedro
authors.initials: JP
authors.lastname: Simas
authors.forename: Colin E
authors.initials: CE
authors.lastname: McVey
mesh_terms: D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D018144:Rhadinovirus / Q000737:chemistry* / Q000502:physiology; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017735:Virus Latency
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins