{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T01:09:20Z","timestamp":1789434560144,"version":"build-2803163510"},"reference-count":39,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2017,4,27]],"date-time":"2017-04-27T00:00:00Z","timestamp":1493251200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Influenza A virus (IAV) infection remains a significant cause of morbidity and mortality worldwide. One key transcription factor that is activated upon IAV infection is nuclear factor Kappa B (NF-\u03baB). NF-\u03baB regulation involves the inhibitor proteins NF-\u03baB inhibitor beta (NFKBIB), (also known as I\u03baB \u03b2), which form complexes with NF-\u03baB to sequester it in the cytoplasm. In this study, microarray data showed differential expression of several microRNAs (miRNAs) on exposure to IAV. Target scan analysis revealed that miR-4776, miR-4514 and miR-4742 potentially target NFKBIB messenger RNA (mRNA). Time-course analysis of primary bronchial epithelial cells (HBEpCs) showed that miR-4776 expression is increased within 1 h of infection, followed by its downregulation 4 h post-exposure to IAV. NFKBIB upregulation of miR-4776 correlated with a decrease in NFKBIB expression within 1 h of infection and a subsequent increase in NFKBIB expression 4 h post-infection. In addition, miRNA ago-immunoprecipitation studies and the three prime untranslated region (3\u2019 UTR) luciferase assay confirmed that miR-4776 targets NFKBIB mRNA. Furthermore, uninfected HBEpCs transfected with miR-4776 mimic showed decreased expression of NFKBIB mRNA. Overexpression of NFKBIB protein in IAV infected cells led to lower levels of IAV. Taken together, our data suggest that miRNA-4776 modulates IAV production in infected cells through NFKBIB expression, possibly through the modulation of NF-\u03baB.<\/jats:p>","DOI":"10.3390\/v9050094","type":"journal-article","created":{"date-parts":[[2017,4,27]],"date-time":"2017-04-27T13:01:15Z","timestamp":1493298075000},"page":"94","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Upregulation of miRNA-4776 in Influenza Virus Infected Bronchial Epithelial Cells Is Associated with Downregulation of NFKBIB and Increased Viral Survival"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6563-611X","authenticated-orcid":false,"given":"Sreekumar","family":"Othumpangat","sequence":"first","affiliation":[{"name":"Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV 26505, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Bryan","sequence":"additional","affiliation":[{"name":"School of Medicine, West Virginia University, Morgantown, WV 26506, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donald","family":"Beezhold","sequence":"additional","affiliation":[{"name":"Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV 26505, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Noti","sequence":"additional","affiliation":[{"name":"Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV 26505, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,4,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1001\/jama.292.11.1333","article-title":"Influenza-associated hospitalizations in the United States","volume":"292","author":"Thompson","year":"2004","journal-title":"JAMA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1101\/gr.082701.108","article-title":"Most mammalian mRNAs are conserved targets of microRNAs","volume":"19","author":"Friedman","year":"2009","journal-title":"Genome Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"8849","DOI":"10.1128\/JVI.00456-10","article-title":"Cellular microRNAs inhibit replication of the H1N1 influenza A virus in infected cells","volume":"84","author":"Song","year":"2010","journal-title":"J. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1038\/sj.cdd.4401838","article-title":"Good cop, bad cop: The different faces of NF-kappaB","volume":"13","author":"Perkins","year":"2006","journal-title":"Cell Death Differ."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/ni.2060","article-title":"Inflammation meets cancer, with NF-kappaB as the matchmaker","volume":"12","author":"Karin","year":"2011","journal-title":"Nat. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1172\/JCI11830","article-title":"NF-kappaB: A key role in inflammatory diseases","volume":"107","author":"Tak","year":"2001","journal-title":"J. Clin. Investig."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2717","DOI":"10.1093\/emboj\/cdg279","article-title":"Caspase 3 activation is essential for efficient influenza virus propagation","volume":"22","author":"Wurzer","year":"2003","journal-title":"EMBO J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"11678","DOI":"10.1074\/jbc.M513286200","article-title":"NFkappaB negatively regulates interferon-induced gene expression and anti-influenza activity","volume":"281","author":"Wei","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2347","DOI":"10.1099\/vir.0.79958-0","article-title":"Active NF-kappaB signalling is a prerequisite for influenza virus infection","volume":"85","author":"Nimmerjahn","year":"2004","journal-title":"J. Gen. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"30931","DOI":"10.1074\/jbc.M403258200","article-title":"NF-\u03baB-dependent induction of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas\/FasL is crucial for efficient influenza virus propagation","volume":"279","author":"Wurzer","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"8307","DOI":"10.1074\/jbc.275.12.8307","article-title":"Influenza virus-induced NF-\u03baB-dependent gene expression is mediated by overexpression of viral proteins and involves oxidative radicals and activation of IkappaB kinase","volume":"275","author":"Flory","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11566","DOI":"10.1128\/JVI.74.24.11566-11573.2000","article-title":"Influenza A virus NS1 protein prevents activation of NF-kappaB and induction of alpha\/beta interferon","volume":"74","author":"Wang","year":"2000","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1111\/j.1462-5822.2005.00678.x","article-title":"Ringing the alarm bells: Signalling and apoptosis in influenza virus infected cells","volume":"8","author":"Ludwig","year":"2006","journal-title":"Cell Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1480","DOI":"10.1128\/jvi.69.3.1480-1484.1995","article-title":"Expression of influenza virus hemagglutinin activates transcription factor NF-\u03baB","volume":"69","author":"Pahl","year":"1995","journal-title":"J. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1111\/cmi.12108","article-title":"The NF-\u03baB inhibitor SC75741 efficiently blocks influenza virus propagation and confers a high barrier for development of viral resistance","volume":"15","author":"Ehrhardt","year":"2013","journal-title":"Cell Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6706","DOI":"10.1038\/sj.onc.1209933","article-title":"Transcriptional regulation via the NF-\u03baB signaling module","volume":"25","author":"Hoffmann","year":"2006","journal-title":"Oncogene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"7068","DOI":"10.1038\/ncomms8068","article-title":"IkappaBbeta enhances the generation of the low-affinity NF-\u03baB\/RelA homodimer","volume":"6","author":"Tsui","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1146\/annurev.immunol.16.1.225","article-title":"NF-\u03baB and Rel proteins: Evolutionarily conserved mediators of immune responses","volume":"16","author":"Ghosh","year":"1998","journal-title":"Annu. Rev. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/S0092-8674(94)90482-0","article-title":"The ubiquitin-proteasome pathway is required for processing the NF-\u03baB1 precursor protein and the activation of NF-kappa B","volume":"78","author":"Palombella","year":"1994","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1007\/s13277-015-3921-1","article-title":"miR-20a enhances cisplatin resistance of human gastric cancer cell line by targeting NFKBIB","volume":"37","author":"Du","year":"2016","journal-title":"Tumour Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1261\/rna.034926.112","article-title":"MicroRNA-182-5p targets a network of genes involved in DNA repair","volume":"19","author":"Krishnan","year":"2013","journal-title":"RNA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.virol.2014.08.007","article-title":"Lung epithelial cells resist influenza A infection by inducing the expression of cytochrome C oxidase VIc which is modulated by miRNA 4276","volume":"468\u2013470","author":"Othumpangat","year":"2014","journal-title":"Virology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.virol.2013.08.031","article-title":"Expression of non-structural-1A binding protein in lung epithelial cells is modulated by miRNA-548an on exposure to influenza A virus","volume":"447","author":"Othumpangat","year":"2013","journal-title":"Virology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3256","DOI":"10.1128\/JCM.40.9.3256-3260.2002","article-title":"Development of a Real-Time Reverse Transcriptase PCR Assay for Type A Influenza Virus and the Avian H5 and H7 Hemagglutinin Subtypes","volume":"40","author":"Spackman","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1146\/annurev-biochem-060308-103103","article-title":"Regulation of mRNA translation and stability by microRNAs","volume":"79","author":"Fabian","year":"2010","journal-title":"Annu. Rev. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.chom.2008.05.002","article-title":"Viral and cellular microRNAs as determinants of viral pathogenesis and immunity","volume":"3","author":"Gottwein","year":"2008","journal-title":"Cell Host Microbe"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/j.virol.2011.01.002","article-title":"Virus-encoded microRNAs","volume":"411","author":"Grundhoff","year":"2011","journal-title":"Virology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1101\/gad.1064703","article-title":"siRNAs can function as miRNAs","volume":"17","author":"Doench","year":"2003","journal-title":"Genes Dev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/S0092-8674(04)00045-5","article-title":"MicroRNAs: Genomics, biogenesis, mechanism, and function","volume":"116","author":"Bartel","year":"2004","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Othumpangat, S., Walton, C., and Piedimonte, G. (2012). MicroRNA-221 modulates RSV replication in human bronchial epithelium by targeting NGF expression. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030030"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1016\/S0092-8674(03)00428-8","article-title":"MicroRNA pathways in flies and worms: Growth, death, fat, stress, and timing","volume":"113","author":"Ambros","year":"2003","journal-title":"Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1038\/nrg1379","article-title":"MicroRNAs: Small RNAs with a big role in gene regulation","volume":"5","author":"He","year":"2004","journal-title":"Nat. Rev. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"9880","DOI":"10.1128\/JVI.00909-08","article-title":"NF-\u03baB signaling differentially regulates influenza virus RNA synthesis","volume":"82","author":"Kumar","year":"2008","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"10736","DOI":"10.1073\/pnas.112338099","article-title":"Cellular transcriptional profiling in influenza A virus-infected lung epithelial cells: The role of the nonstructural NS1 protein in the evasion of the host innate defense and its potential contribution to pandemic influenza","volume":"99","author":"Geiss","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1099\/vir.0.82483-0","article-title":"Effect of the phosphatidylinositol 3-kinase\/Akt pathway on influenza A virus propagation","volume":"88","author":"Shin","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.cell.2005.08.044","article-title":"Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes","volume":"123","author":"Matranga","year":"2005","journal-title":"Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/j.cell.2005.10.022","article-title":"Human RISC couples microRNA biogenesis and posttranscriptional gene silencing","volume":"123","author":"Gregory","year":"2005","journal-title":"Cell"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1007\/978-1-62703-083-0_10","article-title":"Evaluating the microRNA targeting sites by luciferase reporter gene assay","volume":"936","author":"Jin","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/ng1536","article-title":"Combinatorial microRNA target predictions","volume":"37","author":"Krek","year":"2005","journal-title":"Nat. Genet."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/9\/5\/94\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:33:57Z","timestamp":1760207637000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/9\/5\/94"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,4,27]]},"references-count":39,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2017,5]]}},"alternative-id":["v9050094"],"URL":"https:\/\/doi.org\/10.3390\/v9050094","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,4,27]]}}}