Cytomegalovirus cyclin-dependent kinase ortholog vCDK/pUL97 undergoes regulatory interaction with human cyclin H and CDK7 to codetermine viral replication efficiency

Schütz M, Wangen C, Sommerer M, Kögler M, Eickhoff J, Degenhart C, Klebl B, Naing Z, Egilmezer E, Hamilton ST, Rawlinson WD, Sticht H, Marschall M (2023)

Publication Type: Journal article

Publication year: 2023

Journal

Virus Research Elsevier

Book Volume: 335

Article Number: 199200

DOI: 10.1016/j.virusres.2023.199200

Abstract

Human cytomegalovirus (HCMV) infection is shaped by a tightly regulated interplay between viral and cellular proteins. Distinct kinase activities, such as the viral cyclin-dependent kinase ortholog (vCDK) pUL97 and cellular CDK7 are both crucial for efficient viral replication. Previously, we reported that both kinases, vCDK/pUL97 and CDK7, interact with cyclin H, thereby achieving an enhanced level of kinase activity and overall functionality in viral replication. Here we provide a variety of novel results, as generated on a methodologically extended basis, and present a concept for the codetermination of viral replication efficiency through these kinase activities: (i) cyclin H expression, in various human cell types, is substantially upregulated by strains of HCMV including the clinically relevant HCMV Merlin; (ii) vCDK/pUL97 interacts with human cyclin H in both HCMV-infected and plasmid-transfected cell systems; (iii) a doxycycline-inducible shRNA-dependent knock-down (KD) of cyclin H significantly reduces pUL97 activity (qSox in vitro kinase assay); (iv) accordingly, pUL97 in vitro kinase activity is seen significantly increased upon addition of recombinant cyclin H; (v) as a point of specific importance, human CDK7 activity shows an increase by vCDK/pUL97-mediated trans-stimulation (whereas pUL97 is not stimulated by CDK7); (vi) phosphosite-specific antibodies indicate an upregulated CDK7 phosphorylation upon HCMV infection, as mediated through a pUL97-specific modulatory effect (i.e. shown by pUL97 inhibitor treatment or pUL97-deficient viral mutant); (vii) finally, an efficient KD of cyclin H in primary fibroblasts generally results in an impaired HCMV replication efficiency as measured on protein and genomic levels. These results show evidence for the codetermination of viral replication by vCDK/pUL97, cyclin H and CDK7, thus supporting the specific importance of cyclin H as a central regulatory factor, and suggesting novel targeting options for antiviral drugs.

Authors with CRIS profile

Martin Schütz Institute of Clinical and Molecular Virology Markus Sommerer Professur für Kinder- und Jugendpsychiatrie und -psychotherapie Melanie Kögler Institute of Clinical and Molecular Virology Heinrich Sticht Professur für Bioinformatik Manfred Marschall Lehrstuhl für Klinische und Molekulare Virologie

Additional Organisation(s)

Interdisziplinäres Zentrum für Klinische Forschung

Related research project(s)

None - None A088: Regulatory interaction btw. cyclins and cytomegalovirus kinase pUL97: focus on pUL97 drug resistance mutations Feb. 1, 2020 - July 31, 2023

Involved external institutions

Lead Discovery Center GmbH DE Germany (DE) University of New South Wales (UNSW) AU Australia (AU)

How to cite

APA:

Schütz, M., Wangen, C., Sommerer, M., Kögler, M., Eickhoff, J., Degenhart, C.,... Marschall, M. (2023). Cytomegalovirus cyclin-dependent kinase ortholog vCDK/pUL97 undergoes regulatory interaction with human cyclin H and CDK7 to codetermine viral replication efficiency. Virus Research, 335. https://dx.doi.org/10.1016/j.virusres.2023.199200

MLA:

Schütz, Martin, et al. "Cytomegalovirus cyclin-dependent kinase ortholog vCDK/pUL97 undergoes regulatory interaction with human cyclin H and CDK7 to codetermine viral replication efficiency." Virus Research 335 (2023).

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