{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T05:31:09Z","timestamp":1790400669150,"version":"4.1.0"},"reference-count":31,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,8,18]],"date-time":"2021-08-18T00:00:00Z","timestamp":1629244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"maladie infectieuses \u00e9mergentes (ANRS)","award":["ECTZ158023"],"award-info":[{"award-number":["ECTZ158023"]}]},{"name":"maladie infectieuses \u00e9mergentes (ANRS)","award":["ECTZ86820"],"award-info":[{"award-number":["ECTZ86820"]}]},{"DOI":"10.13039\/501100001704","name":"European Society of Clinical Microbiology and Infectious Diseases","doi-asserted-by":"publisher","award":["ESCMID"],"award-info":[{"award-number":["ESCMID"]}],"id":[{"id":"10.13039\/501100001704","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Human cytomegalovirus (HCMV) can cause serious diseases in immunocompromised patients. Current antiviral inhibitors all target the viral DNA polymerase. They have adverse effects, and prolonged treatment can select for drug resistance mutations. Thus, new drugs targeting other stages of replication are an urgent need. The terminase complex (pUL56\u2013pUL89\u2013pUL51) is highly specific, has no counterpart in the human organism, and thus represents a target of choice for new antivirals development. This complex is required for DNA processing and packaging. pUL52 was shown to be essential for the cleavage of concatemeric HCMV DNA and crucial for viral replication, but its functional domains are not yet identified. Polymorphism analysis was performed by sequencing UL52 from 61 HCMV naive strains and from 14 HCMV strains from patients treated with letermovir. Using sequence alignment and homology modeling, we identified conserved regions and potential functional motifs within the pUL52 sequence. Recombinant viruses were generated with specific serine or alanine substitutions in these putative patterns. Within conserved regions, we identified residues essential for viral replication probably involved in CXXC-like or zinc finger motifs. These results suggest that they are essential for pUL52 structure\/function. Thus, these patterns represent potential targets for the development of new antivirals.<\/jats:p>","DOI":"10.3390\/v13081638","type":"journal-article","created":{"date-parts":[[2021,8,18]],"date-time":"2021-08-18T22:51:00Z","timestamp":1629327060000},"page":"1638","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["New Insights into Human Cytomegalovirus pUL52 Structure"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2532-0820","authenticated-orcid":false,"given":"Clotilde","family":"Muller","sequence":"first","affiliation":[{"name":"INSERM, CHU Limoges, University of Limoges, RESINFIT, U1092, F-87000 Limoges, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Alain","sequence":"additional","affiliation":[{"name":"INSERM, CHU Limoges, University of Limoges, RESINFIT, U1092, F-87000 Limoges, France"},{"name":"CHU Limoges, Laboratoire de Bact\u00e9riologie-Virologie-Hygi\u00e8ne, National Reference Center for Herpesviruses (NRCHV), F-87000 Limoges, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7613-437X","authenticated-orcid":false,"given":"Claire","family":"Gourin","sequence":"additional","affiliation":[{"name":"INSERM, CHU Limoges, University of Limoges, RESINFIT, U1092, F-87000 Limoges, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8864-2168","authenticated-orcid":false,"given":"Thomas F.","family":"Baumert","sequence":"additional","affiliation":[{"name":"Institut de Recherche sur les Maladies Virales et H\u00e9patiques, Universit\u00e9 de Strasbourg, 67000 Strasbourg, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1237-1936","authenticated-orcid":false,"given":"Ga\u00ebtan","family":"Ligat","sequence":"additional","affiliation":[{"name":"Institut de Recherche sur les Maladies Virales et H\u00e9patiques, Universit\u00e9 de Strasbourg, 67000 Strasbourg, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2635-0773","authenticated-orcid":false,"given":"S\u00e9bastien","family":"Hantz","sequence":"additional","affiliation":[{"name":"INSERM, CHU Limoges, University of Limoges, RESINFIT, U1092, F-87000 Limoges, France"},{"name":"CHU Limoges, Laboratoire de Bact\u00e9riologie-Virologie-Hygi\u00e8ne, National Reference Center for Herpesviruses (NRCHV), F-87000 Limoges, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1034\/j.1399-3062.2000.020301.x","article-title":"Prevention of Cytomegalovirus Infection in Organ Transplant Recipients","volume":"2","author":"Rubin","year":"2000","journal-title":"Transpl. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.bpobgyn.2016.10.005","article-title":"Fetal Cytomegalovirus Infection","volume":"38","author":"Ville","year":"2017","journal-title":"Best Pract. Res. Clin. Obs. Gynaecol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1128\/CMR.00009-10","article-title":"Antiviral Drug Resistance of Human Cytomegalovirus","volume":"23","author":"Lurain","year":"2010","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2624","DOI":"10.1182\/blood-2016-06-688432","article-title":"How I Treat Resistant Cytomegalovirus Infection in Hematopoietic Cell Transplantation Recipients","volume":"128","author":"Shah","year":"2016","journal-title":"Blood"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.antiviral.2016.02.004","article-title":"Contrasting Effect of New HCMV PUL54 Mutations on Antiviral Drug Susceptibility: Benefits and Limits of 3D Analysis","volume":"129","author":"Andouard","year":"2016","journal-title":"Antivir. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2433","DOI":"10.1056\/NEJMoa1706640","article-title":"Letermovir Prophylaxis for Cytomegalovirus in Hematopoietic-Cell Transplantation","volume":"377","author":"Marty","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Chou, S., Satterwhite, L.E., and Ercolani, R.J. (2018). New Locus of Drug Resistance in the Human Cytomegalovirus UL56 Gene Revealed by In Vitro Exposure to Letermovir and Ganciclovir. Antimicrob. Agents Chemother., 62.","DOI":"10.1128\/AAC.00922-18"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.antiviral.2017.10.019","article-title":"A Third Component of the Human Cytomegalovirus Terminase Complex Is Involved in Letermovir Resistance","volume":"148","author":"Chou","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1093\/femsre\/fuy004","article-title":"The Human Cytomegalovirus Terminase Complex as an Antiviral Target: A Close-up View","volume":"42","author":"Ligat","year":"2018","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Muller, C., Alain, S., Baumert, T.F., Ligat, G., and Hantz, S. (2021). Structures and Divergent Mechanisms in Capsid Maturation and Stabilization Following Genome Packaging of Human Cytomegalovirus and Herpesviruses. Life, 11.","DOI":"10.3390\/life11020150"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1720","DOI":"10.1128\/JVI.01955-12","article-title":"The Human Cytomegalovirus UL51 Protein Is Essential for Viral Genome Cleavage-Packaging and Interacts with the Terminase Subunits PUL56 and PUL89","volume":"87","author":"Borst","year":"2013","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2065","DOI":"10.1128\/JVI.01967-07","article-title":"The Essential Human Cytomegalovirus Gene UL52 Is Required for Cleavage-Packaging of the Viral Genome","volume":"82","author":"Borst","year":"2008","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5860","DOI":"10.1128\/JVI.00384-16","article-title":"The Essential Human Cytomegalovirus Proteins PUL77 and PUL93 Are Structural Components Necessary for Viral Genome Encapsidation","volume":"90","author":"Borst","year":"2016","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"12221","DOI":"10.1128\/JVI.02382-15","article-title":"Human Cytomegalovirus PUL93 Is Required for Viral Genome Cleavage and Packaging","volume":"89","author":"DeRussy","year":"2015","journal-title":"J. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5876","DOI":"10.1128\/JVI.00351-16","article-title":"Intracellular Distribution of Capsid-Associated PUL77 of Human Cytomegalovirus and Interactions with Packaging Proteins and PUL93","volume":"90","author":"Dittmer","year":"2016","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1128\/JVI.01913-14","article-title":"The Putative Herpes Simplex Virus 1 Chaperone Protein UL32 Modulates Disulfide Bond Formation during Infection","volume":"89","author":"Albright","year":"2015","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3938","DOI":"10.1128\/jvi.70.6.3938-3946.1996","article-title":"Properties of the Protein Encoded by the UL32 Open Reading Frame of Herpes Simplex Virus 1","volume":"70","author":"Chang","year":"1996","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2463","DOI":"10.1128\/JVI.72.3.2463-2473.1998","article-title":"The Herpes Simplex Virus Type 1 Cleavage\/Packaging Protein, UL32, Is Involved in Efficient Localization of Capsids to Replication Compartments","volume":"72","author":"Lamberti","year":"1998","journal-title":"J. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.1099\/0022-1317-81-9-2231","article-title":"The Potential Terminase Subunit of Human Cytomegalovirus, PUL56, Is Translocated into the Nucleus by Its Own Nuclear Localization Signal and Interacts with Importin \u03b1","volume":"81","author":"Giesen","year":"2000","journal-title":"J. Gen. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1038\/msb.2011.75","article-title":"Fast, Scalable Generation of High-Quality Protein Multiple Sequence Alignments Using Clustal Omega","volume":"7","author":"Sievers","year":"2011","journal-title":"Mol. Syst. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"W597","DOI":"10.1093\/nar\/gkt376","article-title":"Analysis Tool Web Services from the EMBL-EBI","volume":"41","author":"McWilliam","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"W580","DOI":"10.1093\/nar\/gkv279","article-title":"The EMBL-EBI Bioinformatics Web and Programmatic Tools Framework","volume":"43","author":"Li","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1038\/nprot.2010.5","article-title":"I-TASSER: A Unified Platform for Automated Protein Structure and Function Prediction","volume":"5","author":"Roy","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/nmeth.3213","article-title":"The I-TASSER Suite: Protein Structure and Function Prediction","volume":"12","author":"Yang","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"W174","DOI":"10.1093\/nar\/gkv342","article-title":"I-TASSER Server: New Development for Protein Structure and Function Predictions","volume":"43","author":"Yang","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1128\/JVI.73.10.8320-8329.1999","article-title":"Cloning of the Human Cytomegalovirus (HCMV) Genome as an Infectious Bacterial Artificial Chromosome in Escherichia Coli: A New Approach for Construction of HCMV Mutants","volume":"73","author":"Borst","year":"1999","journal-title":"J. Virol."},{"key":"ref_27","first-page":"125","article-title":"Analysis of the Protein-Coding Content of the Sequence of Human Cytomegalovirus Strain AD169","volume":"154","author":"Chee","year":"1990","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ligat, G., Jacquet, C., Chou, S., Couvreux, A., Alain, S., and Hantz, S. (2017). Identification of a Short Sequence in the HCMV Terminase PUL56 Essential for Interaction with PUL89 Subunit. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-09469-7"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/0022-2836(67)90307-5","article-title":"Selective Extraction of Polyoma DNA from Infected Mouse Cell Cultures","volume":"26","author":"Hirt","year":"1967","journal-title":"J. Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ligat, G., Couvreux, A., Cazal, R., Alain, S., and Hantz, S. (2019). Highlighting of a LAGLIDADG and a Zing Finger Motifs Located in the PUL56 Sequence Crucial for HCMV Replication. Viruses, 11.","DOI":"10.3390\/v11121093"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1177\/135965350801300504","article-title":"Original Article Putative Functional Domains of Human Cytomegalovirus PUL56 Involved in Dimerization and Benzimidazole D-Ribonucleoside Activity","volume":"13","author":"Champier","year":"2008","journal-title":"Antivir. Ther."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/8\/1638\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:46:49Z","timestamp":1760165209000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/8\/1638"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,18]]},"references-count":31,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["v13081638"],"URL":"https:\/\/doi.org\/10.3390\/v13081638","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,18]]}}}