title: Structural genomics of the Epstein-Barr virus.
abstract: Epstein-Barr virus is a herpesvirus that causes infectious mononucleosis, carcinomas and immunoproliferative disease. Its genome encodes 86 proteins, which provided targets for a structural genomics project. After updating the annotation of the genome, 23 open reading frames were chosen for expression in Escherichia coli, initially selecting for those with known enzyme activity and then supplementing this set based on a series of predicted properties, in particular secondary structure. The major obstacle turned out to be poor expression and low solubility. Surprisingly, this could not be overcome by modifications of the constructs, changes of expression temperature or strain or renaturation. Of the eight soluble proteins, five were crystallized using robotic nanolitre-drop crystallization trials, which led to four solved structures. Although these results depended on individual treatment rather than standardized protocols, a high-throughput miniaturized crystallization screening protocol was a key component of success with these difficult proteins.
authors.affiliation: EMBL-Grenoble Outstation, BP 181, Grenoble CEDEX 9, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Tarbouriech
authors.forename: Marlyse
authors.initials: M
authors.lastname: Buisson
authors.forename: Thibault
authors.initials: T
authors.lastname: Géoui
authors.forename: Susan
authors.initials: S
authors.lastname: Daenke
authors.forename: Stephen
authors.initials: S
authors.lastname: Cusack
authors.forename: Wim Pascal
authors.initials: WP
authors.lastname: Burmeister
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D018390:Gene Targeting; D023281:Genomics; D006564:Herpesviridae / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics*; D008958:Models, Molecular; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins