title: A cell free protein fragment complementation assay for monitoring the core interaction of the human cytomegalovirus nuclear egress complex.
abstract: Certain viral protein-protein interactions provide attractive targets for antiviral drug development. Recently, we described a β-lactamase based protein fragment complementation assay (PCA) to study the core interaction of the nuclear egress complex (NEC) of different herpesviruses in cells. Now, to have a cell free assay for inhibitor screens, we expressed split β-lactamase tagged interaction domains of the viral pUL50 and pUL53 proteins representing the NEC of human cytomegalovirus (HCMV) in bacteria. After validation and basic characterization of this NEC-PCA, we tested peptide inhibitors of the pUL50-pUL53 complex. We show that peptides resembling sequences of the first conserved region of pUL53 can inhibit the NEC-PCA. This, on one hand, indicated that the core interaction in the HCMV NEC is mediated by a linear motif. On the other hand it proved that this new pUL50-pUL53 interaction assay allows a simple cell free test for small molecular inhibitors.
authors.affiliation: Max von Pettenkofer Institute, Ludwig-Maximilians University, Gene Center, Feodor Lynen Str. 25, D-81377 Munich, Germany.
authors.forename: Margit
authors.initials: M
authors.lastname: Schnee
authors.forename: Felicia M
authors.initials: FM
authors.lastname: Wagner
authors.forename: Ulrich H
authors.initials: UH
authors.lastname: Koszinowski
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Ruzsics
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D001419:Bacteria; D003587:Cytomegalovirus / Q000502:physiology*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism*; D014773:Virology / Q000379:methods*; D057074:Virus Release; D001618:beta-Lactamases / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D014764:Viral Proteins; C523758:pUL50 protein, cytomegalovirus; C523759:pUL53 protein, cytomegalovirus; D001618:beta-Lactamases