{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T16:22:58Z","timestamp":1792513378795,"version":"4.5.0"},"reference-count":48,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T00:00:00Z","timestamp":1670889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Bayerische Forschungsstiftung","award":["Az-1399-19 DeeP-CMV\/AP-5\/MM"],"award-info":[{"award-number":["Az-1399-19 DeeP-CMV\/AP-5\/MM"]}]},{"name":"Bayerische Forschungsstiftung","award":["5002148\/38726670"],"award-info":[{"award-number":["5002148\/38726670"]}]},{"name":"Bayerische Forschungsstiftung","award":["497739266"],"award-info":[{"award-number":["497739266"]}]},{"name":"Matching Funds Program of the Forschungsstiftung Medizin","award":["Az-1399-19 DeeP-CMV\/AP-5\/MM"],"award-info":[{"award-number":["Az-1399-19 DeeP-CMV\/AP-5\/MM"]}]},{"name":"Matching Funds Program of the Forschungsstiftung Medizin","award":["5002148\/38726670"],"award-info":[{"award-number":["5002148\/38726670"]}]},{"name":"Matching Funds Program of the Forschungsstiftung Medizin","award":["497739266"],"award-info":[{"award-number":["497739266"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["Az-1399-19 DeeP-CMV\/AP-5\/MM"],"award-info":[{"award-number":["Az-1399-19 DeeP-CMV\/AP-5\/MM"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["5002148\/38726670"],"award-info":[{"award-number":["5002148\/38726670"]}]},{"name":"Deutsche Forschungsgemeinschaft","award":["497739266"],"award-info":[{"award-number":["497739266"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Herpesviruses replicate their genomes and assemble their capsids in the host cell nucleus. To progress towards morphogenesis in the cytoplasm, herpesviruses evolved the strategy of nuclear egress as a highly regulated process of nucleo-cytoplasmic capsid transition. The process is conserved among \u03b1-, \u03b2- and \u03b3-herpesviruses and involves the formation of a core and multicomponent nuclear egress complex (NEC). Core NEC is assembled by the interaction between the nucleoplasmic hook protein, i.e., pUL53 (human cytomegalovirus, HCMV), and the integral membrane-associated groove protein, i.e., pUL50. Our study aimed at the question of whether a panherpesviral NEC scaffold may enable hook-into-groove interaction across herpesviral subfamilies. For this purpose, NEC constructs were generated for members of all three subfamilies and analyzed for multi-ligand interaction using a yeast two-hybrid (Y2H) approach with randomized pUL53 mutagenesis libraries. The screening identified ten library clones displaying cross-viral shared hook-into-groove interaction. Interestingly, a slightly modified Y2H screening strategy provided thirteen further changed-hook pUL53 clones having lost parental pUL50 interaction but gained homolog interaction. In addition, we designed a sequence-predicted hybrid construct based on HCMV and Epstein-Barr virus (EBV) core NEC proteins and identified a cross-viral interaction phenotype. Confirmation was provided by applying protein\u2013protein interaction analyses in human cells, such as coimmunoprecipitation settings, confocal nuclear rim colocalization assays, and HCMV \u0394UL53 infection experiments with pUL53-complementing cells. Combined, the study provided the first examples of cross-viral NEC interaction patterns and revealed a higher yield of human cell-confirmed binding clones using a library exchange rate of 3.4 than 2.7. Thus, the study provides improved insights into herpesviral NEC protein binding specificities of core NEC formation. This novel information might be exploited to gain a potential target scaffold for the development of broadly acting NEC-directed inhibitory small molecules.<\/jats:p>","DOI":"10.3390\/cells11244030","type":"journal-article","created":{"date-parts":[[2022,12,14]],"date-time":"2022-12-14T02:54:21Z","timestamp":1670986461000},"page":"4030","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["\u2018Shared-Hook\u2019 and \u2018Changed-Hook\u2019 Binding Activities of Herpesviral Core Nuclear Egress Complexes Identified by Random Mutagenesis"],"prefix":"10.3390","volume":"11","author":[{"given":"Josephine","family":"L\u00f6sing","sequence":"first","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sigrun","family":"H\u00e4ge","sequence":"additional","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Sch\u00fctz","sequence":"additional","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Wagner","sequence":"additional","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Wardin","sequence":"additional","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5644-045X","authenticated-orcid":false,"given":"Heinrich","family":"Sticht","sequence":"additional","affiliation":[{"name":"Division of Bioinformatics, Institute of Biochemistry, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manfred","family":"Marschall","sequence":"additional","affiliation":[{"name":"Institute for Clinical and Molecular Virology, Friedrich-Alexander-Universit\u00e4t Erlangen-N\u00fcrnberg (FAU), 91054 Erlangen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1097\/00005792-198503000-00003","article-title":"Cytomegalovirus infection in the normal host","volume":"64","author":"Cohen","year":"1985","journal-title":"Medicine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1111\/j.1469-0691.2007.01781.x","article-title":"Clinical relevance of cytomegalovirus infection in patients with disorders of the immune system","volume":"13","author":"Steininger","year":"2007","journal-title":"Clin. 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