{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T12:36:08Z","timestamp":1765197368399,"version":"3.46.0"},"posted":{"date-parts":[[2025,12,4]]},"group-title":"Microbiology","reference-count":64,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Herpesviruses and Poxviruses encode secreted chemokine binding proteins that prevent the interaction between chemokines and their cognate G protein coupled receptors to alter chemotactic gradients and intracellular signaling pathways. Human cytomegalovirus (HCMV) encodes the secreted protein UL22A (formerly UL21.5), which is described as a CCL5 (RANTES) binding protein and requires sulfation at two tyrosine residues (Y65 and Y69) for efficient RANTES interaction. In this report, we show that the UL22A protein, and the UL22A Y65 and Y69 residues are necessary for efficient HCMV reactivation from latency in CD34\n                  <jats:sup>+<\/jats:sup>\n                  hematopoietic progenitor cells and that UL22A expression is essential for reactivation in a humanized mouse model of latency. However, RANTES neutralization is not sufficient to complement the\n                  <jats:italic>in vitro<\/jats:italic>\n                  reactivation defect of UL22A mutant viruses. These data suggest that UL22A plays an important role in latency, possibly through interactions with additional chemokines or other types of ligands via its tyrosine residues, in order to mediate efficient HCMV reactivation.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>IMPORTANCE<\/jats:title>\n                  <jats:p>\n                    HCMV is a ubiquitous herpesvirus that infects 60-90% of the population worldwide. In immunocompetent individuals, primary infection is asymptomatic and results in lifelong latent infection in CD34\n                    <jats:sup>+<\/jats:sup>\n                    hematopoietic progenitor cells (HPCs). Viral reactivation remains a major complication for immunosuppressed individuals, but current therapeutics targeting HCMV replication show significant toxicity. Thus, a better understanding of the mechanisms controlling latency and reactivation is necessary to develop new therapeutics targeting these stages of the HCMV lifecycle. We show that virus lacking the HCMV chemokine binding protein UL22A is incapable of efficient reactivation in CD34\n                    <jats:sup>+<\/jats:sup>\n                    HPCs and\n                    <jats:italic>in vivo<\/jats:italic>\n                    . UL22A tyrosine residues important for interaction with the chemokine RANTES are necessary for reactivation. However, neutralizing RANTES does not complement the reactivation defect of UL22A mutant viruses, demonstrating that UL22A has functions other than RANTES binding. Together, our results reveal a novel role for UL22A in HPCs and a new understanding of UL22A-chemokine interactions.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2025.12.04.692306","type":"posted-content","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T12:31:45Z","timestamp":1765197105000},"source":"Crossref","is-referenced-by-count":0,"title":["The human cytomegalovirus chemokine binding protein UL22A is necessary for efficient reactivation from latency in CD34\n                  <sup>+<\/sup>\n                  hematopoietic progenitor cells and humanized mice"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2516-0353","authenticated-orcid":false,"given":"Rebekah L","family":"Turner","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8007-7865","authenticated-orcid":false,"given":"Nicole L","family":"Diggins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luke","family":"Slind","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer","family":"Mitchell","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew H","family":"Pham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher J","family":"Parkins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wilma","family":"Perez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samuel","family":"Medica","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Denton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takeshi F","family":"Andoh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriela M","family":"Webb","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Andrade-Vera","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6828-2492","authenticated-orcid":false,"given":"Daniel N","family":"Streblow","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7579-849X","authenticated-orcid":false,"given":"Patrizia","family":"Caposio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2945-0147","authenticated-orcid":false,"given":"Meaghan H","family":"Hancock","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025120801590791000_2025.12.04.692306v1.1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.14466"},{"key":"2025120801590791000_2025.12.04.692306v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-023-01032-x"},{"key":"2025120801590791000_2025.12.04.692306v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2010.06.004"},{"key":"2025120801590791000_2025.12.04.692306v1.4","doi-asserted-by":"publisher","DOI":"10.1159\/000492161"},{"key":"2025120801590791000_2025.12.04.692306v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cytogfr.2016.02.007"},{"key":"2025120801590791000_2025.12.04.692306v1.6","doi-asserted-by":"crossref","first-page":"e0066425","DOI":"10.1128\/jvi.00664-25","article-title":"Cytomegalovirus latency-the sum of subtleties","volume":"99","year":"2025","journal-title":"J Virol"},{"key":"2025120801590791000_2025.12.04.692306v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-021-00582-z"},{"key":"2025120801590791000_2025.12.04.692306v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiz464"},{"key":"2025120801590791000_2025.12.04.692306v1.9","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.20210"},{"key":"2025120801590791000_2025.12.04.692306v1.10","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1400291"},{"key":"2025120801590791000_2025.12.04.692306v1.11","doi-asserted-by":"crossref","first-page":"e1010355","DOI":"10.1371\/journal.ppat.1010355","article-title":"The non-ELR CXC chemokine encoded by human cytomegalovirus UL146 genotype 5 contains a C-terminal beta-hairpin and induces neutrophil migration as a selective CXCR2 agonist","volume":"18","year":"2022","journal-title":"PLoS Pathog"},{"key":"2025120801590791000_2025.12.04.692306v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)61936-8"},{"key":"2025120801590791000_2025.12.04.692306v1.13","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1995.1814"},{"key":"2025120801590791000_2025.12.04.692306v1.14","doi-asserted-by":"publisher","DOI":"10.1084\/jem.188.5.855"},{"key":"2025120801590791000_2025.12.04.692306v1.15","doi-asserted-by":"crossref","first-page":"6495","DOI":"10.1128\/jvi.71.9.6495-6500.1997","article-title":"Modulation of RANTES production by human cytomegalovirus infection of fibroblasts","volume":"71","year":"1997","journal-title":"J Virol"},{"key":"2025120801590791000_2025.12.04.692306v1.16","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.75.7.3383-3390.2001"},{"key":"2025120801590791000_2025.12.04.692306v1.17","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.73.1.404-410.1999"},{"key":"2025120801590791000_2025.12.04.692306v1.18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002577"},{"key":"2025120801590791000_2025.12.04.692306v1.19","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0504301"},{"key":"2025120801590791000_2025.12.04.692306v1.20","doi-asserted-by":"publisher","DOI":"10.1186\/s12964-016-0154-x"},{"key":"2025120801590791000_2025.12.04.692306v1.21","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-08-372516"},{"key":"2025120801590791000_2025.12.04.692306v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2011.06.009"},{"key":"2025120801590791000_2025.12.04.692306v1.23","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0407233101"},{"key":"2025120801590791000_2025.12.04.692306v1.24","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2022.102232"},{"key":"2025120801590791000_2025.12.04.692306v1.25","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M407536200"},{"key":"2025120801590791000_2025.12.04.692306v1.26","doi-asserted-by":"crossref","first-page":"8490","DOI":"10.1002\/anie.201703059","article-title":"Sulfation of the Human Cytomegalovirus Protein UL22A Enhances Binding to the Chemokine RANTES","volume":"56","year":"2017","journal-title":"Angew Chem Int Ed Engl"},{"key":"2025120801590791000_2025.12.04.692306v1.27","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.67.9.5502-5513.1993"},{"key":"2025120801590791000_2025.12.04.692306v1.28","first-page":"185","article-title":"Human cytomegalovirus UL21.5 gene is expressed as an \u201cearly-late\u201d gene in cultured human fibroblasts","volume":"45","year":"2001","journal-title":"Acta Virol"},{"key":"2025120801590791000_2025.12.04.692306v1.29","doi-asserted-by":"crossref","unstructured":"Shnayder M , Nachshon A , Krishna B , Poole E , Boshkov A , Binyamin A , Maza I , Sinclair J , Schwartz M , Stern-Ginossar N . 2018. Defining the Transcriptional Landscape during Cytomegalovirus Latency with Single-Cell RNA Sequencing. mBio 9.","DOI":"10.1128\/mBio.00013-18"},{"key":"2025120801590791000_2025.12.04.692306v1.30","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1710522114"},{"key":"2025120801590791000_2025.12.04.692306v1.31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.252630899"},{"key":"2025120801590791000_2025.12.04.692306v1.32","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-788-4_7"},{"key":"2025120801590791000_2025.12.04.692306v1.33","doi-asserted-by":"crossref","unstructured":"Crawford LB , Hancock MH , Struthers HM , Streblow DN , Yurochko AD , Caposio P , Goodrum FD , Nelson JA . 2021. CD34(+) Hematopoietic Progenitor Cell Subsets Exhibit Differential Ability To Maintain Human Cytomegalovirus Latency and Persistence. J Virol 95.","DOI":"10.1128\/JVI.02105-20"},{"key":"2025120801590791000_2025.12.04.692306v1.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2009.06.008"},{"key":"2025120801590791000_2025.12.04.692306v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2015.06.006"},{"key":"2025120801590791000_2025.12.04.692306v1.36","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2010.08.001"},{"key":"2025120801590791000_2025.12.04.692306v1.37","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.006231"},{"key":"2025120801590791000_2025.12.04.692306v1.38","doi-asserted-by":"publisher","DOI":"10.4149\/av_2015_04_360"},{"key":"2025120801590791000_2025.12.04.692306v1.39","doi-asserted-by":"publisher","DOI":"10.1084\/jem.191.3.573"},{"key":"2025120801590791000_2025.12.04.692306v1.40","doi-asserted-by":"publisher","DOI":"10.1126\/science.1455233"},{"key":"2025120801590791000_2025.12.04.692306v1.41","doi-asserted-by":"crossref","first-page":"4356","DOI":"10.1128\/jvi.71.6.4356-4363.1997","article-title":"The purified myxoma virus gamma interferon receptor homolog M-T7 interacts with the heparin-binding domains of chemokines","volume":"71","year":"1997","journal-title":"J Virol"},{"key":"2025120801590791000_2025.12.04.692306v1.42","doi-asserted-by":"publisher","DOI":"10.3390\/molecules20022138"},{"key":"2025120801590791000_2025.12.04.692306v1.43","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1182\/blood.V94.1.62.413k04_62_73","article-title":"Human CD34(+) cells express CXCR4 and its ligand stromal cell-derived factor-1. Implications for infection by T-cell tropic human immunodeficiency virus","volume":"94","year":"1999","journal-title":"Blood"},{"key":"2025120801590791000_2025.12.04.692306v1.44","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2003-07-2607"},{"key":"2025120801590791000_2025.12.04.692306v1.45","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20011284"},{"key":"2025120801590791000_2025.12.04.692306v1.46","doi-asserted-by":"publisher","DOI":"10.1634\/stemcells.18-5-374"},{"key":"2025120801590791000_2025.12.04.692306v1.47","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.cyto.2018.01.021","article-title":"Regulation of hematopoiesis by the chemokine system","volume":"109","year":"2018","journal-title":"Cytokine"},{"key":"2025120801590791000_2025.12.04.692306v1.48","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-11-391730"},{"key":"2025120801590791000_2025.12.04.692306v1.49","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1182\/blood.V88.4.1277.bloodjournal8841277","article-title":"Direct infection of CD34+ progenitor cells by human cytomegalovirus: evidence for inhibition of hematopoiesis and viral replication","volume":"88","year":"1996","journal-title":"Blood"},{"key":"2025120801590791000_2025.12.04.692306v1.50","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V88.12.4526.bloodjournal88124526"},{"key":"2025120801590791000_2025.12.04.692306v1.51","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.75.16.7543-7554.2001"},{"key":"2025120801590791000_2025.12.04.692306v1.52","doi-asserted-by":"crossref","first-page":"1421","DOI":"10.3389\/fmicb.2019.01421","article-title":"Chemokine-Binding Proteins Encoded by Parapoxvirus of Red Deer of New Zealand Display Evidence of Gene Duplication and Divergence of Ligand Specificity","volume":"10","year":"2019","journal-title":"Front Microbiol"},{"key":"2025120801590791000_2025.12.04.692306v1.53","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2009.01425.x"},{"key":"2025120801590791000_2025.12.04.692306v1.54","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.009589-0"},{"key":"2025120801590791000_2025.12.04.692306v1.55","doi-asserted-by":"publisher","DOI":"10.1016\/j.cyto.2008.05.016"},{"key":"2025120801590791000_2025.12.04.692306v1.56","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.177.8.5567"},{"key":"2025120801590791000_2025.12.04.692306v1.57","doi-asserted-by":"publisher","DOI":"10.1097\/01.TP.0000131173.52424.84"},{"key":"2025120801590791000_2025.12.04.692306v1.58","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.170.3.1435"},{"key":"2025120801590791000_2025.12.04.692306v1.59","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.1.624-630.2003"},{"key":"2025120801590791000_2025.12.04.692306v1.60","doi-asserted-by":"publisher","DOI":"10.1172\/JCI200214358"},{"key":"2025120801590791000_2025.12.04.692306v1.61","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001321"},{"key":"2025120801590791000_2025.12.04.692306v1.62","doi-asserted-by":"publisher","DOI":"10.1084\/jem.194.3.301"},{"key":"2025120801590791000_2025.12.04.692306v1.63","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1008666"},{"key":"2025120801590791000_2025.12.04.692306v1.64","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.83286-0"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2025.12.04.692306","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T12:31:47Z","timestamp":1765197107000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2025.12.04.692306"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,4]]},"references-count":64,"URL":"https:\/\/doi.org\/10.64898\/2025.12.04.692306","relation":{},"subject":[],"published":{"date-parts":[[2025,12,4]]},"subtype":"preprint"}}