title: New insight into the phosphorylation-regulated intranuclear localization of human cytomegalovirus pUL69 mediated by cyclin-dependent kinases (CDKs) and viral CDK orthologue pUL97.
abstract: Cyclin-dependent kinases (CDKs) are multifaceted regulators involved in the replication of human cytomegalovirus. Recently, we demonstrated an interaction of CDK9-cyclin T1 as well as viral CDK orthologue pUL97 with the viral regulator pUL69, thereby leading to pUL69-activating phosphorylation. Here, we demonstrate that colocalization and direct pUL69-cyclin T1 interaction is independent of viral strains and host cell types. In vitro phosphorylation of pUL69 by CDK9 or pUL97 did not occur in a single site-specific manner, but at multiple sites. The previously described fine-speckled nuclear aggregation of pUL69 was assigned to the late phase of viral replication. CDK inhibitors, including a novel inhibitor of the CDK-activating kinase CDK7, massively intensified this fine-speckled accumulation. Interestingly, we also observed spontaneous pUL69 accumulation in the absence of inhibitors at a lower frequency. These findings provide new insight into pUL69 kinase interregulation and emphasize the importance of pUL69 phosphorylation for correct intranuclear localization.
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Laura
authors.initials: L
authors.lastname: Graf
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Feichtinger
authors.affiliation: Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
authors.forename: Zin
authors.initials: Z
authors.lastname: Naing
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Corina
authors.initials: C
authors.lastname: Hutterer
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Milbradt
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Rike
authors.initials: R
authors.lastname: Webel
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Wagner
authors.affiliation: Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Scott
authors.affiliation: Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
authors.forename: Stuart T
authors.initials: ST
authors.lastname: Hamilton
authors.affiliation: Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, Sydney, Australia.
authors.forename: William D
authors.initials: WD
authors.lastname: Rawlinson
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stamminger
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Thomas
authors.affiliation: Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Marschall
mesh_terms: D042863:Cyclin-Dependent Kinase 9 / Q000378:metabolism*; D003587:Cytomegalovirus / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D015534:Trans-Activators / Q000378:metabolism*
chemical_list: D015534:Trans-Activators; C446019:pUL69 protein, Human herpesvirus 5; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; C467969:CDK9 protein, human; D042863:Cyclin-Dependent Kinase 9