title: Proteomic Interaction Patterns between Human Cyclins, the Cyclin-Dependent Kinase Ortholog pUL97 and Additional Cytomegalovirus Proteins.
abstract: The human cytomegalovirus (HCMV)-encoded cyclin-dependent kinase (CDK) ortholog pUL97 associates with human cyclin B1 and other types of cyclins. Here, the question was addressed whether cyclin interaction of pUL97 and additional viral proteins is detectable by mass spectrometry-based approaches. Proteomic data were validated by coimmunoprecipitation (CoIP), Western blot, in vitro kinase and bioinformatic analyses. Our findings suggest that: (i) pUL97 shows differential affinities to human cyclins; (ii) pUL97 inhibitor maribavir (MBV) disrupts the interaction with cyclin B1, but not with other cyclin types; (iii) cyclin H is identified as a new high-affinity interactor of pUL97 in HCMV-infected cells; (iv) even more viral phosphoproteins, including all known substrates of pUL97, are detectable in the cyclin-associated complexes; and (v) a first functional validation of pUL97-cyclin B1 interaction, analyzed by in vitro kinase assay, points to a cyclin-mediated modulation of pUL97 substrate preference. In addition, our bioinformatic analyses suggest individual, cyclin-specific binding interfaces for pUL97-cyclin interaction, which could explain the different strengths of interactions and the selective inhibitory effect of MBV on pUL97-cyclin B1 interaction. Combined, the detection of cyclin-associated proteins in HCMV-infected cells suggests a complex pattern of substrate phosphorylation and a role of cyclins in the fine-modulation of pUL97 activities.
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. mirjam.steingruber@viro.med.uni-erlangen.de.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Steingruber
authors.affiliation: Laboratoire Biologie à Grande Echelle (BGE), Exploring the Dynamics of Proteomes (EDyP), Université Grenoble Alpes, iRTSV-BGE, 38000 Grenoble, France. alexandra.kraut@cea.fr.|CEA, iRTSV-BGE, Grenoble 38000, France. alexandra.kraut@cea.fr.|INSERM, BGE, Grenoble 38000, France. alexandra.kraut@cea.fr.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Kraut
authors.affiliation: Division of Bioinformatics, Institute of Biochemistry, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. eileen.socher@fau.de.
authors.forename: Eileen
authors.initials: E
authors.lastname: Socher
authors.affiliation: Division of Bioinformatics, Institute of Biochemistry, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. heinrich.sticht@fau.de.
authors.forename: Heinrich
authors.initials: H
authors.lastname: Sticht
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. anna.reichel@viro.med.uni-erlangen.de.
authors.forename: Anna
authors.initials: A
authors.lastname: Reichel
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. thomas.stamminger@viro.med.uni-erlangen.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stamminger
authors.affiliation: Division of Biochemistry, Department of Biology, Friedrich-Alexander University of Erlangen-Nürnberg, 91058 Erlangen, Germany. bushra.amin@fau.de.
authors.forename: Bushra
authors.initials: B
authors.lastname: Amin
authors.affiliation: Laboratoire Biologie à Grande Echelle (BGE), Exploring the Dynamics of Proteomes (EDyP), Université Grenoble Alpes, iRTSV-BGE, 38000 Grenoble, France. yohann.coute@cea.fr.|CEA, iRTSV-BGE, Grenoble 38000, France. yohann.coute@cea.fr.|INSERM, BGE, Grenoble 38000, France. yohann.coute@cea.fr.
authors.forename: Yohann
authors.initials: Y
authors.lastname: Couté
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. corina.hutterer@viro.med.uni-erlangen.de.
authors.forename: Corina
authors.initials: C
authors.lastname: Hutterer
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, 91054 Erlangen, Germany. manfred.marschall@viro.med.uni-erlangen.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Marschall
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D019295:Computational Biology; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism*; D003587:Cytomegalovirus / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D040901:Proteomics; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D016213:Cyclins; C436448:UL89 protein, Cytomegalovirus; D014764:Viral Proteins; D018844:Cyclin-Dependent Kinases
keywords: CDK ortholog pUL97; cyclin-associated protein complexes; human cyclins; human cytomegalovirus; molecular modeling; proteomic analysis