Highly Conserved Interaction Profiles between Clinically Relevant Mutants of the Cytomegalovirus CDK-like Kinase pUL97 and Human Cyclins: Functional Significance of Cyclin H

Schütz M, Müller R, Socher E, Wangen C, Full F, Wyler E, Wong DD, Scherer M, Stamminger T, Chou S, Rawlinson WD, Hamilton ST, Sticht H, Marschall M (2022)

Publication Type: Journal article

Publication year: 2022

Journal

Original Authors: Martin Schütz, Regina Müller, Eileen Socher, Christina Wangen, Florian Full, Emanuel Wyler, Diana Wong, Myriam Scherer, Thomas Stamminger, Sunwen Chou, William D. Rawlinson, Stuart T. Hamilton, Heinrich Sticht, Manfred Marschall

Book Volume: 23

Pages Range: 11814

Issue: 19

DOI: 10.3390/ijms231911814

Abstract

The complex host interaction network of human cytomegalovirus (HCMV) involves the regulatory protein kinase pUL97, which represents a viral cyclin-dependent kinase (CDK) ortholog. pUL97 interacts with the three human cyclin types T1, H, and B1, whereby the binding region of cyclin T1 and the pUL97 oligomerization region were both assigned to amino acids 231-280. We further addressed the question of whether HCMVs harboring mutations in ORF-UL97, i.e., short deletions or resistance-conferring point mutations, are affected in the interaction with human cyclins and viral replication. To this end, clinically relevant UL97 drug-resistance-conferring mutants were analyzed by whole-genome sequencing and used for genetic marker transfer experiments. The recombinant HCMVs indicated conservation of pUL97–cyclin interaction, since all viral UL97 point mutants continued to interact with the analyzed cyclin types and exerted wild-type-like replication fitness. In comparison, recombinant HCMVs UL97 Δ231-280 and also the smaler deletion Δ236-275, but not Δ241-270, lost interaction with cyclins T1 and H, showed impaired replication efficiency, and also exhibited reduced kinase activity. Moreover, a cellular knock-out of cyclins B1 or T1 did not alter HCMV replication phenotypes or pUL97 kinase activity, possibly indicating alternative, compensatory pUL97–cyclin interactions. In contrast, however, cyclin H knock-out, similar to virus deletion mutants in the pUL97–cyclin H binding region, exhibited strong defective phenotypes of HCMV replication, as supported by reduced pUL97 kinase activity in a cyclin H-dependent coexpression setting. Thus, cyclin H proved to be a very relevant determinant of pUL97 kinase activity and viral replication efficiency. As a conclusion, the results provide evidence for the functional importance of pUL97–cyclin interaction. High selective pressure on the formation of pUL97–cyclin complexes was identified by the use of clinically relevant mutants

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University of New South Wales (UNSW) Australia (AU) Universitätsklinikum Ulm Germany (DE) Albert-Ludwigs-Universität Freiburg Germany (DE) Prince of Wales Hospital (POWH) Australia (AU) Universität Ulm Germany (DE) Oregon Health and Science University (OSHU) United States (USA) (US) Max-Delbrück-Centrum für Molekulare Medizin / Max Delbrück Center for Molecular Medicine (MDC) Berlin-Buch Germany (DE)

How to cite

APA:

Schütz, M., Müller, R., Socher, E., Wangen, C., Full, F., Wyler, E.,... Marschall, M. (2022). Highly Conserved Interaction Profiles between Clinically Relevant Mutants of the Cytomegalovirus CDK-like Kinase pUL97 and Human Cyclins: Functional Significance of Cyclin H. International Journal of Molecular Sciences, 23, 11814. https://dx.doi.org/10.3390/ijms231911814

MLA:

Schütz, Martin, et al. "Highly Conserved Interaction Profiles between Clinically Relevant Mutants of the Cytomegalovirus CDK-like Kinase pUL97 and Human Cyclins: Functional Significance of Cyclin H." International Journal of Molecular Sciences 23 (2022): 11814.

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