{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:27:37Z","timestamp":1760059657195,"version":"build-2065373602"},"reference-count":103,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T00:00:00Z","timestamp":1751414400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["148361"],"award-info":[{"award-number":["148361"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Infections with cytomegalovirus (CMV) can result in increased morbidity and mortality in immunocompromised patients. The pUL97 kinase is a critical enzyme in the regulation of CMV replication. Although it does not phosphorylate deoxynucleosides, this enzyme is involved in the first phosphorylation step of ganciclovir (GCV), a viral DNA polymerase inhibitor. In contrast, maribavir (MBV) is a specific inhibitor of pUL97 kinase activity. In this paper, we analyzed the already-reported amino acid changes, conferring resistance to MBV and cross-resistance to GCV, in the pUL97 protein structure, predicted with AlphaFold2. Docking experiments suggest that MBV is a dual-site inhibitor, targeting ATP binding and substrate phosphorylation. Substitutions that confer resistance to MBV only may directly or indirectly alter the shape of the cavity in the vicinity of the invariant K355 in the putative ATP binding site, without affecting the viral growth. The most frequently encountered T409M substitution may correspond to a gatekeeper mutation. Substitutions that induce cross-resistance to MBV and GCV may directly or indirectly affect the environment of D456 and N461 residues in the catalytic loop, with reduced viral replicative capacity. These results have implications for the clinical use of MBV as well as for the design of novel pUL97 kinase inhibitors.<\/jats:p>","DOI":"10.3390\/v17070941","type":"journal-article","created":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T07:55:43Z","timestamp":1751442943000},"page":"941","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In Silico Analysis of Mechanisms of Maribavir-Induced Inhibition and Drug Resistance Mutations in pUL97 Kinase Structural Prediction with AlphaFold2"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4102-0094","authenticated-orcid":false,"given":"Jocelyne","family":"Piret","sequence":"first","affiliation":[{"name":"Research Center of the CHU de Quebec-Laval University, Quebec City, QC G1V 4G2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3027-4662","authenticated-orcid":false,"given":"Guy","family":"Boivin","sequence":"additional","affiliation":[{"name":"Research Center of the CHU de Quebec-Laval University, Quebec City, QC G1V 4G2, Canada"},{"name":"Department of Pediatrics, Faculty of Medicine, Laval University, Quebec City, QC G1V 0A6, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,2]]},"reference":[{"key":"ref_1","unstructured":"Boivin, G., and Limaye, A.P. (2023). Cytomegalovirus. Goldman-Cecil Medicine, Elsevier."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-981-16-0267-2_1","article-title":"Antiviral Drugs Against Herpesviruses","volume":"1322","author":"Piret","year":"2021","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"65","DOI":"10.3390\/idr16010005","article-title":"Management of Cytomegalovirus Infections in the Era of the Novel Antiviral Players, Letermovir and Maribavir","volume":"16","author":"Piret","year":"2024","journal-title":"Infect. Dis. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1128\/AAC.49.3.1039-1045.2005","article-title":"In vitro activity and mechanism of action of methylenecyclopropane analogs of nucleosides against herpesvirus replication","volume":"49","author":"Kern","year":"2005","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4682","DOI":"10.1128\/AAC.00571-11","article-title":"Cyclopropavir inhibits the normal function of the human cytomegalovirus UL97 kinase","volume":"55","author":"James","year":"2011","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1006\/viro.1997.8523","article-title":"The human cytomegalovirus UL97 protein is phosphorylated and a component of virions","volume":"231","author":"Fairhurst","year":"1997","journal-title":"Virology"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"6340","DOI":"10.1128\/jvi.70.9.6340-6346.1996","article-title":"The UL97 gene product of human cytomegalovirus is an early-late protein with a nuclear localization but is not a nucleoside kinase","volume":"70","author":"Michel","year":"1996","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1099\/vir.0.026799-0","article-title":"Two isoforms of the protein kinase pUL97 of human cytomegalovirus are differentially regulated in their nuclear translocation","volume":"92","author":"Webel","year":"2011","journal-title":"J. Gen. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1756","DOI":"10.1099\/vir.0.040592-0","article-title":"Nuclear import of isoforms of the cytomegalovirus kinase pUL97 is mediated by differential activity of NLS1 and NLS2 both acting through classical importin-alpha binding","volume":"93","author":"Webel","year":"2012","journal-title":"J. Gen. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1128\/JVI.74.2.975-986.2000","article-title":"Accumulation of virion tegument and envelope proteins in a stable cytoplasmic compartment during human cytomegalovirus replication: Characterization of a potential site of virus assembly","volume":"74","author":"Sanchez","year":"2000","journal-title":"J. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4776","DOI":"10.1128\/JVI.00192-14","article-title":"Differential properties of cytomegalovirus pUL97 kinase isoforms affect viral replication and maribavir susceptibility","volume":"88","author":"Webel","year":"2014","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1128\/jvi.71.1.405-411.1997","article-title":"The human cytomegalovirus UL97 protein is a protein kinase that autophosphorylates on serines and threonines","volume":"71","author":"He","year":"1997","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11943","DOI":"10.1128\/JVI.76.23.11943-11952.2002","article-title":"Relationship between autophosphorylation and phosphorylation of exogenous substrates by the human cytomegalovirus UL97 protein kinase","volume":"76","author":"Baek","year":"2002","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2006","DOI":"10.1002\/pmic.201400607","article-title":"Human cytomegalovirus pUL97 kinase induces global changes in the infected cell phosphoproteome","volume":"15","author":"Oberstein","year":"2015","journal-title":"Proteomics"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1002\/rmv.615","article-title":"Function of human cytomegalovirus UL97 kinase in viral infection and its inhibition by maribavir","volume":"19","author":"Prichard","year":"2009","journal-title":"Rev. Med. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/S0968-0004(97)01144-4","article-title":"The cyclin box fold: Protein recognition in cell-cycle and transcription control","volume":"22","author":"Noble","year":"1997","journal-title":"Trends Biochem. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1126\/science.1152095","article-title":"Phosphorylation of retinoblastoma protein by viral protein with cyclin-dependent kinase function","volume":"320","author":"Hume","year":"2008","journal-title":"Science"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5054","DOI":"10.1128\/JVI.02174-07","article-title":"Human cytomegalovirus UL97 kinase activity is required for the hyperphosphorylation of retinoblastoma protein and inhibits the formation of nuclear aggresomes","volume":"82","author":"Prichard","year":"2008","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.virol.2017.09.009","article-title":"Phosphorylation of transcriptional regulators in the retinoblastoma protein pathway by UL97, the viral cyclin-dependent kinase encoded by human cytomegalovirus","volume":"512","author":"Iwahori","year":"2017","journal-title":"Virology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6583","DOI":"10.1074\/jbc.M116.773150","article-title":"Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins","volume":"292","author":"Iwahori","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11988","DOI":"10.1128\/JVI.78.21.11988-12011.2004","article-title":"Global analysis of host cell gene expression late during cytomegalovirus infection reveals extensive dysregulation of cell cycle gene expression and induction of Pseudomitosis independent of US28 function","volume":"78","author":"Hertel","year":"2004","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3213","DOI":"10.3390\/v5123213","article-title":"The cyclin-dependent kinase ortholog pUL97 of human cytomegalovirus interacts with cyclins","volume":"5","author":"Graf","year":"2013","journal-title":"Viruses"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4582","DOI":"10.3390\/v7082834","article-title":"The Interaction between Cyclin B1 and Cytomegalovirus Protein Kinase pUL97 is Determined by an Active Kinase Domain","volume":"7","author":"Steingruber","year":"2015","journal-title":"Viruses"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Steingruber, M., Kraut, A., Socher, E., Sticht, H., Reichel, A., Stamminger, T., Amin, B., Coute, Y., Hutterer, C., and Marschall, M. (2016). Proteomic Interaction Patterns between Human Cyclins, the Cyclin-Dependent Kinase Ortholog pUL97 and Additional Cytomegalovirus Proteins. Viruses, 8.","DOI":"10.3390\/v8080219"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6188","DOI":"10.1074\/jbc.RA118.007049","article-title":"Cyclins B1, T1, and H differ in their molecular mode of interaction with cytomegalovirus protein kinase pUL97","volume":"294","author":"Steingruber","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Schutz, M., Steingruber, M., Socher, E., Muller, R., Wagner, S., Kogel, M., Sticht, H., and Marschall, M. (2021). Functional Relevance of the Interaction between Human Cyclins and the Cytomegalovirus-Encoded CDK-Like Protein Kinase pUL97. Viruses, 13.","DOI":"10.3390\/v13071248"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4456","DOI":"10.1128\/JVI.73.5.4456-4460.1999","article-title":"Cellular elongation factor 1delta is modified in cells infected with representative alpha-, beta-, or gammaherpesviruses","volume":"73","author":"Kawaguchi","year":"1999","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.virol.2004.03.015","article-title":"Phosphorylation of the RNA polymerase II carboxyl-terminal domain in human cytomegalovirus-infected cells and in vitro by the viral UL97 protein kinase","volume":"324","author":"Baek","year":"2004","journal-title":"Virology"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Hamirally, S., Kamil, J.P., Ndassa-Colday, Y.M., Lin, A.J., Jahng, W.J., Baek, M.C., Noton, S., Silva, L.A., Simpson-Holley, M., and Knipe, D.M. (2009). Viral mimicry of Cdc2\/cyclin-dependent kinase 1 mediates disruption of nuclear lamina during human cytomegalovirus nuclear egress. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000275"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5019","DOI":"10.1128\/JVI.00007-13","article-title":"Inactivation of retinoblastoma protein does not overcome the requirement for human cytomegalovirus UL97 in lamina disruption and nuclear egress","volume":"87","author":"Reim","year":"2013","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1128\/JVI.02358-13","article-title":"Human cytomegalovirus UL50 and UL53 recruit viral protein kinase UL97, not protein kinase C, for disruption of nuclear lamina and nuclear egress in infected cells","volume":"88","author":"Sharma","year":"2014","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1038\/358160a0","article-title":"Human cytomegalovirus UL97 open reading frame encodes a protein that phosphorylates the antiviral ganciclovir","volume":"358","author":"Littler","year":"1992","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1038\/358162a0","article-title":"A protein kinase homologue controls phosphorylation of ganciclovir in human cytomegalovirus-infected cells","volume":"358","author":"Sullivan","year":"1992","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2473","DOI":"10.1073\/pnas.82.8.2473","article-title":"Metabolic activation of the nucleoside analog 9-[(2-hydroxy-1-(hydroxymethyl)ethoxy]methyl)guanine in human diploid fibroblasts infected with human cytomegalovirus","volume":"82","author":"Biron","year":"1985","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1128\/jvi.53.3.776-780.1985","article-title":"Inhibition of cellular DNA polymerase alpha and human cytomegalovirus-induced DNA polymerase by the triphosphates of 9-(2-hydroxyethoxymethyl)guanine and 9-(1,3-dihydroxy-2-propoxymethyl)guanine","volume":"53","author":"Mar","year":"1985","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2365","DOI":"10.1128\/AAC.46.8.2365-2372.2002","article-title":"Potent and selective inhibition of human cytomegalovirus replication by 1263W94, a benzimidazole L-riboside with a unique mode of action","volume":"46","author":"Biron","year":"2002","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1128\/JVI.77.2.905-914.2003","article-title":"The human cytomegalovirus UL97 protein kinase, an antiviral drug target, is required at the stage of nuclear egress","volume":"77","author":"Krosky","year":"2003","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/2042-4280-1-4","article-title":"The effects of maribavir on the autophosphorylation of ganciclovir resistant mutants of the cytomegalovirus UL97 protein","volume":"1","author":"Emery","year":"2010","journal-title":"Herpesviridae"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3470","DOI":"10.1128\/AAC.00577-06","article-title":"Maribavir antagonizes the antiviral action of ganciclovir on human cytomegalovirus","volume":"50","author":"Chou","year":"2006","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3093","DOI":"10.1128\/AAC.00468-10","article-title":"Stereoselective phosphorylation of cyclopropavir by pUL97 and competitive inhibition by maribavir","volume":"54","author":"Gentry","year":"2010","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1007\/s40265-022-01677-4","article-title":"Maribavir: First Approval","volume":"82","author":"Kang","year":"2022","journal-title":"Drugs"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5.8.1","DOI":"10.1002\/0471250953.bi0508s52","article-title":"Protein Structure and Function Prediction Using I-TASSER","volume":"52","author":"Yang","year":"2015","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1038\/s41592-020-0848-2","article-title":"Macromolecular modeling and design in Rosetta: Recent methods and frameworks","volume":"17","author":"Leman","year":"2020","journal-title":"Nat. Methods"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly accurate protein structure prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1038\/s41592-022-01488-1","article-title":"ColabFold: Making protein folding accessible to all","volume":"19","author":"Mirdita","year":"2022","journal-title":"Nat. Methods"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of biomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Evans, R., O\u2019Neill, M., Pritzel, A., Antropova, N., Senior, A., Green, T., \u017d\u00eddek, A., Bates, R., Blackwell, S., and Yim, J. (2021). Protein complex prediction with AlphaFold-Multimer. bioRxiv.","DOI":"10.1101\/2021.10.04.463034"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2722","DOI":"10.1093\/bioinformatics\/btt473","article-title":"lDDT: A local superposition-free score for comparing protein structures and models using distance difference tests","volume":"29","author":"Mariani","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"W431","DOI":"10.1093\/nar\/gkab314","article-title":"Mol* Viewer: Modern web app for 3D visualization and analysis of large biomolecular structures","volume":"49","author":"Sehnal","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"W404","DOI":"10.1093\/nar\/gkad350","article-title":"PAE viewer: A webserver for the interactive visualization of the predicted aligned error for multimer structure predictions and crosslinks","volume":"51","author":"Elfmann","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1002\/pro.3330","article-title":"MolProbity: More and better reference data for improved all-atom structure validation","volume":"27","author":"Williams","year":"2018","journal-title":"Protein Sci."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1002\/prot.10286","article-title":"Structure validation by Calpha geometry: Phi, psi and Cbeta deviation","volume":"50","author":"Lovell","year":"2003","journal-title":"Proteins"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1021\/jm0306430","article-title":"Glide: A new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy","volume":"47","author":"Friesner","year":"2004","journal-title":"J. Med. Chem."},{"key":"ref_54","unstructured":"Schrodinger, K.K. (The PyMOL Molecular Graphics System, 2010). The PyMOL Molecular Graphics System, version 1.3rl."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"W296","DOI":"10.1093\/nar\/gky427","article-title":"SWISS-MODEL: Homology modelling of protein structures and complexes","volume":"46","author":"Waterhouse","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"7044","DOI":"10.1021\/jm060696s","article-title":"Analysis of the structure-activity relationship of four herpesviral UL97 subfamily protein kinases reveals partial but not full functional conservation","volume":"49","author":"Romaker","year":"2006","journal-title":"J. Med. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1126\/science.3291115","article-title":"The protein kinase family: Conserved features and deduced phylogeny of the catalytic domains","volume":"241","author":"Hanks","year":"1988","journal-title":"Science"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1099\/0022-1317-70-5-1151","article-title":"Alpha-, beta- and gammaherpesviruses encode a putative phosphotransferase","volume":"70","author":"Chee","year":"1989","journal-title":"J. Gen. Virol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.bbapap.2007.08.009","article-title":"Conserved herpesvirus protein kinases","volume":"1784","author":"Gershburg","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/S0092-8674(00)81092-2","article-title":"Active and inactive protein kinases: Structural basis for regulation","volume":"85","author":"Johnson","year":"1996","journal-title":"Cell"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1439","DOI":"10.1099\/0022-1317-82-6-1439","article-title":"Inhibitors of human cytomegalovirus replication drastically reduce the activity of the viral protein kinase pUL97","volume":"82","author":"Marschall","year":"2001","journal-title":"J. Gen. Virol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1038\/363595a0","article-title":"Crystal structure of cyclin-dependent kinase 2","volume":"363","author":"Rosenblatt","year":"1993","journal-title":"Nature"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1038\/376313a0","article-title":"Mechanism of CDK activation revealed by the structure of a cyclinA-CDK2 complex","volume":"376","author":"Jeffrey","year":"1995","journal-title":"Nature"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1038\/nsb0896-696","article-title":"Structural basis of cyclin-dependent kinase activation by phosphorylation","volume":"3","author":"Russo","year":"1996","journal-title":"Nat. Struct. Biol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"8746","DOI":"10.1074\/jbc.274.13.8746","article-title":"Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2 structure and activity","volume":"274","author":"Brown","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1074\/jbc.M007337200","article-title":"Kinetic basis for activation of CDK2\/cyclin A by phosphorylation","volume":"276","author":"Hagopian","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1128\/JVI.01787-07","article-title":"Accelerated evolution of maribavir resistance in a cytomegalovirus exonuclease domain II mutant","volume":"82","author":"Chou","year":"2008","journal-title":"J. Virol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"104616","DOI":"10.1016\/j.antiviral.2019.104616","article-title":"Novel UL97 drug resistance mutations identified at baseline in a clinical trial of maribavir for resistant or refractory cytomegalovirus infection","volume":"172","author":"Chou","year":"2019","journal-title":"Antivir. Res."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1002\/rmv.574","article-title":"Cytomegalovirus UL97 mutations in the era of ganciclovir and maribavir","volume":"18","author":"Chou","year":"2008","journal-title":"Rev. Med. Virol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.drup.2018.01.003","article-title":"Thymidine kinase and protein kinase in drug-resistant herpesviruses: Heads of a Lernaean Hydra","volume":"37","author":"Topalis","year":"2018","journal-title":"Drug Resist. Updat."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"29289","DOI":"10.1074\/jbc.M504096200","article-title":"Structural basis for autoinhibition and mutational activation of eukaryotic initiation factor 2alpha protein kinase GCN2","volume":"280","author":"Padyana","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1021\/jm990401t","article-title":"Binding mode of the 4-anilinoquinazoline class of protein kinase inhibitor: X-ray crystallographic studies of 4-anilinoquinazolines bound to cyclin-dependent kinase 2 and p38 kinase","volume":"43","author":"Shewchuk","year":"2000","journal-title":"J. Med. Chem."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"4540","DOI":"10.1021\/jm960402a","article-title":"High-resolution crystal structures of human cyclin-dependent kinase 2 with and without ATP: Bound waters and natural ligand as guides for inhibitor design","volume":"39","author":"Kim","year":"1996","journal-title":"J. Med. Chem."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1016\/j.jmb.2004.04.043","article-title":"Structural modes of stabilization of permissive phosphorylation sites in protein kinases: Distinct strategies in Ser\/Thr and Tyr kinases","volume":"339","author":"Krupa","year":"2004","journal-title":"J. Mol. Biol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"18626","DOI":"10.1016\/S0021-9258(17)46675-0","article-title":"Identification of phosphorylation sites in the recombinant catalytic subunit of cAMP-dependent protein kinase","volume":"268","author":"Yonemoto","year":"1993","journal-title":"J. Biol. Chem."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"2154","DOI":"10.1021\/bi00060a005","article-title":"Crystal structure of the catalytic subunit of cAMP-dependent protein kinase complexed with MgATP and peptide inhibitor","volume":"32","author":"Zheng","year":"1993","journal-title":"Biochemistry"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"17783","DOI":"10.1073\/pnas.0607656103","article-title":"Surface comparison of active and inactive protein kinases identifies a conserved activation mechanism","volume":"103","author":"Kornev","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1002\/j.1460-2075.1993.tb05725.x","article-title":"Phosphotransferase and substrate binding mechanism of the cAMP-dependent protein kinase catalytic subunit from porcine heart as deduced from the 2.0 A structure of the complex with Mn2+ adenylyl imidodiphosphate and inhibitor peptide PKI(5-24)","volume":"12","author":"Bossemeyer","year":"1993","journal-title":"EMBO J."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.antiviral.2016.08.005","article-title":"The chemical class of quinazoline compounds provides a core structure for the design of anticytomegaloviral kinase inhibitors","volume":"134","author":"Hutterer","year":"2016","journal-title":"Antivir. Res."},{"key":"ref_80","doi-asserted-by":"crossref","unstructured":"Marschall, M., Schutz, M., Wild, M., Socher, E., Wangen, C., Dhotre, K., Rawlinson, W.D., and Sticht, H. (2024). Understanding the Cytomegalovirus Cyclin-Dependent Kinase Ortholog pUL97 as a Multifaceted Regulator and an Antiviral Drug Target. Cells, 13.","DOI":"10.3390\/cells13161338"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1021\/cb5008376","article-title":"Small molecule substrate phosphorylation site inhibitors of protein kinases: Approaches and challenges","volume":"10","author":"Breen","year":"2015","journal-title":"ACS Chem. Biol."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"15494","DOI":"10.1128\/JVI.79.24.15494-15502.2005","article-title":"Human cytomegalovirus UL97 Kinase is required for the normal intranuclear distribution of pp65 and virion morphogenesis","volume":"79","author":"Prichard","year":"2005","journal-title":"J. Virol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"10659","DOI":"10.1128\/JVI.00497-07","article-title":"Human cytomegalovirus protein kinase UL97 forms a complex with the tegument phosphoprotein pp65","volume":"81","author":"Kamil","year":"2007","journal-title":"J. Virol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1128\/JVI.02426-14","article-title":"Human cytomegalovirus UL97 phosphorylates the viral nuclear egress complex","volume":"89","author":"Sharma","year":"2015","journal-title":"J. Virol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"15625","DOI":"10.1021\/bi0268910","article-title":"Specificity determinants of recruitment peptides bound to phospho-CDK2\/cyclin A","volume":"41","author":"Lowe","year":"2002","journal-title":"Biochemistry"},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1093\/protein\/gzx068","article-title":"Short linear motif core and flanking regions modulate retinoblastoma protein binding affinity and specificity","volume":"31","author":"Palopoli","year":"2018","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1038\/36038","article-title":"Structure of the retinoblastoma tumour-suppressor pocket domain bound to a peptide from HPV E7","volume":"391","author":"Lee","year":"1998","journal-title":"Nature"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/1743-422X-6-9","article-title":"Conserved retinoblastoma protein-binding motif in human cytomegalovirus UL97 kinase minimally impacts viral replication but affects susceptibility to maribavir","volume":"6","author":"Gill","year":"2009","journal-title":"Virol. J."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"13130","DOI":"10.1039\/D4SC01088D","article-title":"The prospect of substrate-based kinase inhibitors to improve target selectivity and overcome drug resistance","volume":"15","author":"Biswas","year":"2024","journal-title":"Chem. Sci."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"1349","DOI":"10.1080\/17460441.2023.2265303","article-title":"Advances in protein kinase drug discovery through targeting gatekeeper mutations","volume":"18","author":"Guo","year":"2023","journal-title":"Expert. Opin. Drug Discov."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1086\/518514","article-title":"Cytomegalovirus UL97 kinase mutations that confer maribavir resistance","volume":"196","author":"Chou","year":"2007","journal-title":"J. Infect. Dis."},{"key":"ref_92","doi-asserted-by":"crossref","unstructured":"Schutz, M., Muller, R., Socher, E., Wangen, C., Full, F., Wyler, E., Wong, D., Scherer, M., Stamminger, T., and Chou, S. (2022). Highly Conserved Interaction Profiles between Clinically Relevant Mutants of the Cytomegalovirus CDK-like Kinase pUL97 and Human Cyclins: Functional Significance of Cyclin H. Int. J. Mol. Sci., 23.","DOI":"10.21203\/rs.3.rs-1676959\/v1"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.antiviral.2012.05.013","article-title":"Effects on maribavir susceptibility of cytomegalovirus UL97 kinase ATP binding region mutations detected after drug exposure in vitro and in vivo","volume":"95","author":"Chou","year":"2012","journal-title":"Antivir. Res."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"3375","DOI":"10.1128\/AAC.00511-13","article-title":"Cytomegalovirus UL97 kinase catalytic domain mutations that confer multidrug resistance","volume":"57","author":"Chou","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1093\/infdis\/jiab029","article-title":"Phenotype and Genotype Study of Novel C480F Maribavir-Ganciclovir Cross-Resistance Mutation Detected in Hematopoietic Stem Cell and Solid Organ Transplant Recipients","volume":"224","author":"Plault","year":"2021","journal-title":"J. Infect. Dis."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1128\/AAC.01726-13","article-title":"Human cytomegalovirus UL97 kinase is involved in the mechanism of action of methylenecyclopropane analogs with 6-ether and -thioether substitutions","volume":"58","author":"Chou","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.jcv.2010.08.008","article-title":"Contrasting effects on ganciclovir susceptibility and replicative capacity of two mutations at codon 466 of the human cytomegalovirus UL97 gene","volume":"49","author":"Martin","year":"2010","journal-title":"J. Clin. Virol. Off. Publ. Pan Am. Soc. Clin. Virol."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"105139","DOI":"10.1016\/j.antiviral.2021.105139","article-title":"Ganciclovir and maribavir susceptibility phenotypes of cytomegalovirus UL97 ATP binding region mutations detected by expanded genotypic testing","volume":"193","author":"Chou","year":"2021","journal-title":"Antivir. Res."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"105792","DOI":"10.1016\/j.antiviral.2023.105792","article-title":"Ganciclovir and maribavir cross-resistance revisited: Relative drug susceptibilities of canonical cytomegalovirus mutants","volume":"222","author":"Chou","year":"2024","journal-title":"Antivir. Res."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1086\/338362","article-title":"Cytomegalovirus UL97 phosphotransferase mutations that affect susceptibility to ganciclovir","volume":"185","author":"Chou","year":"2002","journal-title":"J. Infect. Dis."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1093\/infdis\/jiaa462","article-title":"Drug Resistance Mutations and Associated Phenotypes Detected in Clinical Trials of Maribavir for Treatment of Cytomegalovirus Infection","volume":"226","author":"Chou","year":"2022","journal-title":"J. Infect. Dis."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1042\/BST20210837","article-title":"Regulatory spine RS3 residue of protein kinases: A lipophilic bystander or a decisive element in the small-molecule kinase inhibitor binding?","volume":"50","author":"Shevchenko","year":"2022","journal-title":"Biochem. Soc. Trans."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.antiviral.2019.01.011","article-title":"Clinical development of letermovir and maribavir: Overview of human cytomegalovirus drug resistance","volume":"163","author":"Piret","year":"2019","journal-title":"Antivir. Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/7\/941\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:03:01Z","timestamp":1760032981000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/17\/7\/941"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,2]]},"references-count":103,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["v17070941"],"URL":"https:\/\/doi.org\/10.3390\/v17070941","relation":{},"ISSN":["1999-4915"],"issn-type":[{"type":"electronic","value":"1999-4915"}],"subject":[],"published":{"date-parts":[[2025,7,2]]}}}