{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T22:13:31Z","timestamp":1762640011877,"version":"build-2065373602"},"reference-count":51,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,20]],"date-time":"2020-10-20T00:00:00Z","timestamp":1603152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P01 NS065719"],"award-info":[{"award-number":["P01 NS065719"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Polyomaviruses are small, non-enveloped DNA tumor viruses that cause serious disease in immunosuppressed people, including progressive multifocal leukoencephalopathy (PML) in patients infected with JC polyomavirus, but the molecular events mediating polyomavirus entry are poorly understood. Through genetic knockdown approaches, we identified phosphoinositide 3\u2032-kinase \u03b3 (PI3K\u03b3) and its regulatory subunit PIK3R5 as cellular proteins that facilitate infection of human SVG-A glial cells by JCPyV. PI3K\u03b1 appears less important for polyomavirus infection than PI3K\u03b3. CRISPR\/Cas9-mediated knockout of PIK3R5 or PI3K\u03b3 inhibited infection by authentic JCPyV and by JC pseudovirus. PI3K\u03b3 knockout also inhibited infection by BK and Merkel Cell pseudoviruses, other pathogenic human polyomaviruses, and SV40, an important model polyomavirus. Reintroduction of the wild-type PI3K\u03b3 gene into the PI3K\u03b3 knock-out SVG-A cells rescued the JCPyV infection defect. Disruption of the PI3K\u03b3 pathway did not block binding of JCPyV to cells or virus internalization, implying that PI3K\u03b3 facilitates some intracellular step(s) of infection. These results imply that agents that inhibit PI3K\u03b3 signaling may have a role in managing polyomavirus infections.<\/jats:p>","DOI":"10.3390\/v12101190","type":"journal-article","created":{"date-parts":[[2020,10,20]],"date-time":"2020-10-20T09:28:23Z","timestamp":1603186103000},"page":"1190","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Phosphoinositide 3\u2032-Kinase \u03b3 Facilitates Polyomavirus Infection"],"prefix":"10.3390","volume":"12","author":[{"given":"Paul","family":"Clark","sequence":"first","affiliation":[{"name":"Department of Genetics, Yale School of Medicine, P.O. Box 208005, New Haven, CT 06520-8005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gretchen V.","family":"Gee","sequence":"additional","affiliation":[{"name":"Molecular Biology, Cell Biology and Biochemistry, Brown University, Box G-E431, Lab of Molecular Medicine, 70 Ship Street, Providence, RI 02912, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brandon S.","family":"Albright","sequence":"additional","affiliation":[{"name":"Department of Genetics, Yale School of Medicine, P.O. 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Box 208024, New Haven, CT 06520-8024, USA"},{"name":"Yale Cancer Center, P.O. Box 208028, New Haven, CT 06520-8028, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1515\/hsz-2016-0345","article-title":"The biology of JC polyomavirus","volume":"398","author":"Assetta","year":"2017","journal-title":"Biol. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1146\/annurev-virology-101416-041439","article-title":"Progressive multifocal Leukoencephalopathy: Endemic viruses and lethal brain disease","volume":"4","author":"Haley","year":"2017","journal-title":"Annu. Rev. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1016\/S1473-3099(09)70226-9","article-title":"Progressive multifocal leukoencephalopathy in HIV-1 infection","volume":"9","author":"Cinque","year":"2009","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1128\/CMR.05031-11","article-title":"Molecular biology, epidemiology, and pathogenesis of progressive multifocal Leukoencephalopathy, the JC virus-induced demyelinating disease of the human brain","volume":"25","author":"Ferenczy","year":"2012","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"458","DOI":"10.1128\/jvi.51.2.458-469.1984","article-title":"Human polyomavirus JC virus genome","volume":"51","author":"Frisque","year":"1984","journal-title":"J. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1056\/NEJMp0906248","article-title":"Reemergence of PML in natalizumab-treated patients--new cases, same concerns","volume":"361","author":"Major","year":"2009","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1056\/NEJMc055235","article-title":"Natalizumab and progressive multifocal Leukoencephalopathy","volume":"353","author":"Adelman","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1007\/s13365-016-0427-6","article-title":"Reassessing the risk of natalizumab-associated PML","volume":"22","author":"Berger","year":"2016","journal-title":"J. NeuroVirology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e235","DOI":"10.1212\/NXI.0000000000000235","article-title":"A link between long-term natalizumab dosing in MS and PML: Putting the puzzle together","volume":"3","author":"Major","year":"2016","journal-title":"Neurol. Neuroimmunol. Neuroinflammation"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1007\/s13365-015-0392-5","article-title":"2nd International Conference on Progressive Multifocal Leukoencephalopathy (PML) 2015: JCV virology, progressive multifocal leukoencephalopathy pathogenesis, diagnosis and risk stratification, and new approaches to prevention and treatment","volume":"21","author":"Patera","year":"2015","journal-title":"J. NeuroVirology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1212\/CON.0000000000000245","article-title":"Central nervous system infections with Immunomodulatory therapies","volume":"21","author":"Williamson","year":"2015","journal-title":"Contin. Lifelong Learn. Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1007\/s40263-015-0226-2","article-title":"Infection risk in patients on multiple sclerosis therapeutics","volume":"29","author":"Williamson","year":"2015","journal-title":"CNS Drugs"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1056\/NEJM198607243150405","article-title":"Association of BK Viruria with hemorrhagic cystitis in recipients of bone marrow transplants","volume":"315","author":"Arthur","year":"1986","journal-title":"N. Engl. J. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1253","DOI":"10.1016\/S0140-6736(71)91776-4","article-title":"New human Papovavirus (B.K.) isolated from urine after renal transplantation","volume":"297","author":"Gardner","year":"1971","journal-title":"Lancet"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"374","DOI":"10.2215\/CJN.02021205","article-title":"Histologic versus molecular diagnosis of BK polyomavirus-associated nephropathy: A shifting paradigm?","volume":"1","author":"Drachenberg","year":"2006","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_16","first-page":"25","article-title":"Polyomavirus BK nephropathy: A (Re-)emerging complication in renal transplantation","volume":"2","author":"Hirsch","year":"2002","journal-title":"Arab. Archaeol. Epigr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1093\/ndt\/15.3.324","article-title":"BK-virus nephropathy in renal transplants\u2014Tubular necrosis, MHC-class II expression and rejection in a puzzling game","volume":"15","author":"Nickeleit","year":"2000","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1111\/j.1399-3062.2010.00567.x","article-title":"JC virus-associated nephropathy in a renal transplant recipient and comparative analysis of previous cases","volume":"13","author":"Kantarci","year":"2010","journal-title":"Transpl. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1097\/01.TP.0000090749.42791.14","article-title":"The first case of JC virus allograft nephropathy","volume":"76","author":"Kazory","year":"2003","journal-title":"Transplantation"},{"key":"ref_20","first-page":"2887","article-title":"A case of primary JC Polyomavirus infection-associated nephropathy","volume":"14","author":"Lautenschlager","year":"2014","journal-title":"Arab. Archaeol. Epigr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1002\/jmv.20037","article-title":"Association of JC virus with tubulointerstitial nephritis in a renal allograft recipient","volume":"72","author":"Wen","year":"2004","journal-title":"J. Med. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1681\/ASN.2010080877","article-title":"Inhibitory interactions between BK and JC virus among kidney transplant recipients","volume":"22","author":"Cheng","year":"2011","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1097\/01.tp.0000269706.59977.a5","article-title":"Polyomavirus BK versus JC replication and nephropathy in renal transplant recipients: A prospective evaluation","volume":"84","author":"Drachenberg","year":"2007","journal-title":"Transplantation"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1126\/science.1152586","article-title":"Clonal Integration of a Polyomavirus in human merkel cell carcinoma","volume":"319","author":"Feng","year":"2008","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/j.chom.2010.05.006","article-title":"Merkel cell Polyomavirus and two previously unknown Polyomaviruses are chronically shed from human skin","volume":"7","author":"Schowalter","year":"2010","journal-title":"Cell Host Microbe"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"16272","DOI":"10.1073\/pnas.0806526105","article-title":"T antigen mutations are a human tumor-specific signature for Merkel cell polyomavirus","volume":"105","author":"Shuda","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Berrios, C., Padi, M., Keibler, M.A., Park, D.E., Molla, V., Cheng, J., Lee, S.M., Stephanopoulos, G., Quackenbush, J., and DeCaprio, J.A. (2016). Merkel cell Polyomavirus small T antigen promotes pro-glycolytic metabolic perturbations required for transformation. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1006020"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6118","DOI":"10.1128\/JVI.00385-13","article-title":"Merkel cell Polyomavirus large T antigen has growth-promoting and inhibitory activities","volume":"87","author":"Cheng","year":"2013","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1073\/pnas.1818798116","article-title":"Dual inhibition of MDM2 and MDM4 in virus-positive Merkel cell carcinoma enhances the p53 response","volume":"116","author":"Park","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"An, P., Robles, M.T.S., Duray, A.M., Cantalupo, P.G., and Pipas, J.M. (2019). Human polyomavirus BKV infection of endothelial cells results in interferon pathway induction and persistence. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1007505"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e00903-16","DOI":"10.1128\/mBio.00903-16","article-title":"JC Polyomavirus infection of primary human renal epithelial cells is controlled by a type I IFN-induced response","volume":"7","author":"Assetta","year":"2016","journal-title":"mBio"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0969-2126(96)00020-2","article-title":"The structure of simian virus 40 refined at 3.1 \u00e5 resolution","volume":"4","author":"Stehle","year":"1996","journal-title":"Structure"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1016\/j.cell.2007.09.038","article-title":"Simian virus 40 depends on ER protein folding and quality control factors for entry into host cells","volume":"131","author":"Schelhaas","year":"2007","journal-title":"Cell"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Chen, Y.-J., Liu, X., and Tsai, B. (2019). SV40 hijacks cellular transport, membrane penetration, and disassembly machineries to promote infection. Viruses, 11.","DOI":"10.3390\/v11100917"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1038\/ncb2339","article-title":"BAP31 and BiP are essential for dislocation of SV40 from the endoplasmic reticulum to the cytosol","volume":"13","author":"Geiger","year":"2011","journal-title":"Nat. Cell Biol."},{"key":"ref_36","first-page":"459","article-title":"Oncocytic tumors of the kidney. 11 cases (1978\u20131987)","volume":"23","author":"Brucher","year":"1989","journal-title":"Ann. d\u2019Urologie"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e00101-11","DOI":"10.1128\/mBio.00101-11","article-title":"BiP and multiple DNAJ molecular chaperones in the endoplasmic reticulum are required for efficient simian virus 40 infection","volume":"2","author":"Goodwin","year":"2011","journal-title":"mBio"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1111\/tra.12649","article-title":"Towards a molecular understanding of endosomal trafficking by Retromer and Retriever","volume":"20","author":"Chen","year":"2019","journal-title":"Traffic"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e00729-13","DOI":"10.1128\/mBio.00729-13","article-title":"A retrograde trafficking inhibitor of ricin and shiga-like toxins inhibits infection of cells by human and monkey Polyomaviruses","volume":"4","author":"Nelson","year":"2013","journal-title":"mBio"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Dushane, J.K., and Maginnis, M.S. (2019). Human DNA virus exploitation of the MAPK-ERK cascade. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20143427"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Dushane, J.K., Mayberry, C.L., Wilczek, M.P., Nichols, S.L., and Maginnis, M.S. (2019). JCPyV-Induced MAPK signaling activates transcription factors during infection. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20194779"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"01529-17","DOI":"10.1128\/JVI.01529-17","article-title":"ERK is a critical regulator of JC Polyomavirus infection","volume":"92","author":"Dushane","year":"2018","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2590","DOI":"10.1016\/j.jmb.2018.06.024","article-title":"Virus\u2013receptor interactions: The key to cellular invasion","volume":"430","author":"Maginnis","year":"2018","journal-title":"J. Mol. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1210","DOI":"10.1073\/pnas.84.5.1210","article-title":"Regulation of c-myc and c-fos mRNA levels by polyomavirus: Distinct roles for the capsid protein VP1 and the viral early proteins","volume":"84","author":"Zullo","year":"1987","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1016\/0042-6822(89)90425-X","article-title":"Extension of JC virus host range to monkey cells by insertion of a simian virus 40 enhancer into the JC virus regulatory region","volume":"170","author":"Vacante","year":"1989","journal-title":"Virology"},{"key":"ref_46","first-page":"9","article-title":"Propagation and assay of the JC virus","volume":"165","author":"Liu","year":"2001","journal-title":"Methods Mol. Boil. (Clifton N.J.)"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Luo, Y., Motamedi, N., Magaldi, T.G., Gee, G.V., Atwood, W.J., and DiMaio, D. (2016). Interaction between Simian virus 40 major capsid protein VP1 and cell surface Ganglioside GM1 triggers vacuole formation. mBio, 7.","DOI":"10.1128\/mBio.00297-16"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.virusres.2013.09.030","article-title":"Pseudovirus mimics cell entry and trafficking of the human polyomavirus JCPyV","volume":"178","author":"Gee","year":"2013","journal-title":"Virus Res."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Pastrana, D.V., Tolstov, Y.L., Becker, J.C., Moore, P.S., Chang, Y., and Buck, C.B. (2009). Quantitation of human Seroresponsiveness to merkel cell Polyomavirus. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000578"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1038\/nmeth.3047","article-title":"Improved vectors and genome-wide libraries for CRISPR screening","volume":"11","author":"Sanjana","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/S0092-8674(00)80187-7","article-title":"The G\u03b2\u03b3 sensitivity of a PI3K is dependent upon a tightly associated adaptor, p101","volume":"89","author":"Stephens","year":"1997","journal-title":"Cell"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/10\/1190\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:24:40Z","timestamp":1760178280000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/12\/10\/1190"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,20]]},"references-count":51,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["v12101190"],"URL":"https:\/\/doi.org\/10.3390\/v12101190","relation":{},"ISSN":["1999-4915"],"issn-type":[{"type":"electronic","value":"1999-4915"}],"subject":[],"published":{"date-parts":[[2020,10,20]]}}}