{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T07:40:29Z","timestamp":1789371629347,"version":"build-2803163510"},"reference-count":44,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2016,5,24]],"date-time":"2016-05-24T00:00:00Z","timestamp":1464048000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01 AI087643-01A1"],"award-info":[{"award-number":["R01 AI087643-01A1"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"name":"The James W. McLaughlin Endowment","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"The Sealy Center for Vaccine Development, UTMB","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm\u2019-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn\/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn\/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn\/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment.<\/jats:p>","DOI":"10.3390\/v8060151","type":"journal-article","created":{"date-parts":[[2016,5,24]],"date-time":"2016-05-24T21:53:40Z","timestamp":1464126820000},"page":"151","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression"],"prefix":"10.3390","volume":"8","author":[{"given":"Inaia","family":"Phoenix","sequence":"first","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nandadeva","family":"Lokugamage","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shoko","family":"Nishiyama","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8318-2783","authenticated-orcid":false,"given":"Tetsuro","family":"Ikegami","sequence":"additional","affiliation":[{"name":"Department of Pathology, The University of Texas Medical Branch, Galveston, TX 77555, USA"},{"name":"The Sealy Center for Vaccine Development, The University of Texas Medical Branch, Galveston, TX 77555, USA"},{"name":"The Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch, Galveston, TX 77555, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2016,5,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Pepin, M., Bouloy, M., Bird, B.H., Kemp, A., and Paweska, J. (2010). Rift Valley fever virus (Bunyaviridae: Phlebovirus): An update on pathogenesis, molecular epidemiology, vectors, diagnostics and prevention. Vet. Res., 41.","DOI":"10.1051\/vetres\/2010033"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.antiviral.2012.06.001","article-title":"Molecular biology and genetic diversity of Rift Valley fever virus","volume":"95","author":"Ikegami","year":"2012","journal-title":"Antivir. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"493","DOI":"10.3390\/v3050493","article-title":"The pathogenesis of Rift Valley fever","volume":"3","author":"Ikegami","year":"2011","journal-title":"Viruses"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"883","DOI":"10.2460\/javma.234.7.883","article-title":"Rift Valley fever virus","volume":"234","author":"Bird","year":"2009","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1017\/S0022172400062434","article-title":"Rift Valley fever virus (family Bunyaviridae, genus Phlebovirus). Isolations from Diptera collected during an inter-epizootic period in Kenya","volume":"95","author":"Linthicum","year":"1985","journal-title":"J. Hyg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"D69","DOI":"10.1016\/j.vaccine.2009.07.046","article-title":"Rift valley fever vaccines","volume":"27","author":"Ikegami","year":"2009","journal-title":"Vaccine"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"9798","DOI":"10.1128\/JVI.78.18.9798-9806.2004","article-title":"NSs protein of Rift Valley fever virus blocks interferon production by inhibiting host gene transcription","volume":"78","author":"Billecocq","year":"2004","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1371","DOI":"10.1128\/JVI.75.3.1371-1377.2001","article-title":"Genetic evidence for an interferon-antagonistic function of Rift Valley fever virus nonstructural protein NSs","volume":"75","author":"Bouloy","year":"2001","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/S0092-8674(04)00132-1","article-title":"TFIIH transcription factor, a target for the Rift Valley hemorrhagic fever virus","volume":"116","author":"Dubaele","year":"2004","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Le May, N., Mansuroglu, Z., Leger, P., Josse, T., Blot, G., Billecocq, A., Flick, R., Jacob, Y., Bonnefoy, E., and Bouloy, M. (2008). A SAP30 complex inhibits IFN-beta expression in Rift Valley fever virus infected cells. PLoS Pathog., 4.","DOI":"10.1371\/journal.ppat.0040013"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Ikegami, T., Narayanan, K., Won, S., Kamitani, W., Peters, C.J., and Makino, S. (2009). Rift Valley fever virus NSs protein promotes post-transcriptional downregulation of protein kinase PKR and inhibits eIF2\u03b1 phosphorylation. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000287"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6234","DOI":"10.1128\/JVI.02255-10","article-title":"NSs protein of Rift Valley fever virus promotes post-translational downregulation of the TFIIH subunit p62","volume":"85","author":"Kalveram","year":"2011","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4365","DOI":"10.1128\/JVI.02148-08","article-title":"NSs protein of Rift Valley fever virus induces the specific degradation of the double-stranded RNA-dependent protein kinase","volume":"83","author":"Habjan","year":"2009","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/0042-6822(89)90442-X","article-title":"Rift Valley fever virus M segment: Phlebovirus expression strategy and protein glycosylation","volume":"170","author":"Kakach","year":"1989","journal-title":"Virology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1016\/0042-6822(88)90562-4","article-title":"Rift Valley fever virus M segment: Cell-free transcription and translation of virus-complementary RNA","volume":"164","author":"Suzich","year":"1988","journal-title":"Virology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1128\/jvi.64.4.1549-1555.1990","article-title":"Expression strategy of a phlebovirus: Biogenesis of proteins from the Rift Valley fever virus M segment","volume":"64","author":"Suzich","year":"1990","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1083\/jcb.115.4.887","article-title":"An analysis of vertebrate mRNA sequences: Intimations of translational control","volume":"115","author":"Kozak","year":"1991","journal-title":"J. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kreher, F., Tamietti, C., Gommet, C., Guillemot, L., Ermonval, M., Failloux, A.B., Panthier, J.J., Bouloy, M., and Flamand, M. (2014). The Rift Valley fever accessory proteins NSm and P78\/NSm-Gn are determinants of virus propagation in vertebrate and invertebrate hosts. Emerg. Microb. Infect., 3.","DOI":"10.1038\/emi.2014.71"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/0264-410X(91)90314-V","article-title":"Further evaluation of a mutagen-attenuated Rift Valley fever vaccine in sheep","volume":"9","author":"Morrill","year":"1991","journal-title":"Vaccine"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.2460\/ajvr.1987.48.07.1042","article-title":"Pathogenicity and immunogenicity of a mutagen-attenuated Rift Valley fever virus immunogen in pregnant ewes","volume":"48","author":"Morrill","year":"1987","journal-title":"Am. J. Vet. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1110","DOI":"10.2460\/ajvr.1997.58.10.1110","article-title":"Safety of a mutagen-attenuated Rift Valley fever virus vaccine in fetal and neonatal bovids","volume":"58","author":"Morrill","year":"1997","journal-title":"Am. J. Vet. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7262","DOI":"10.1128\/JVI.00135-15","article-title":"Rift Valley fever virus MP-12 vaccine is fully attenuated by a combination of partial attenuations in the S, M, and L segments","volume":"89","author":"Ikegami","year":"2015","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3735","DOI":"10.1128\/JVI.02241-15","article-title":"The L-, M- and S-segments of Rift Valley fever virus MP-12 vaccine independently contribute to a temperature-sensitive phenotype","volume":"90","author":"Nishiyama","year":"2016","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1128\/CVI.00114-15","article-title":"Evaluation of the efficacy, potential for vector transmission, and duration of immunity of MP-12, an attenuated Rift Valley fever virus vaccine candidate, in sheep","volume":"22","author":"Miller","year":"2015","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8274","DOI":"10.1128\/JVI.00476-06","article-title":"NSm and 78-kilodalton proteins of Rift Valley fever virus are nonessential for viral replication in cell culture","volume":"80","author":"Won","year":"2006","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/j.virol.2006.09.035","article-title":"The NSm proteins of Rift Valley fever virus are dispensable for maturation, replication and infection","volume":"359","author":"Gerrard","year":"2007","journal-title":"Virology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1128\/JVI.02192-12","article-title":"The C-terminal region of Rift Valley fever virus NSm protein targets the protein to the mitochondrial outer membrane and exerts anti-apoptotic function","volume":"87","author":"Terasaki","year":"2013","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.virol.2006.08.002","article-title":"Synthesis, proteolytic processing and complex formation of N-terminally nested precursor proteins of the Rift Valley fever virus glycoproteins","volume":"357","author":"Gerrard","year":"2007","journal-title":"Virology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/j.ymthe.2004.10.016","article-title":"Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo","volume":"11","author":"Tannous","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"12200","DOI":"10.1128\/JVI.76.23.12200-12210.2002","article-title":"Characterization of the Golgi retention motif of Rift Valley fever virus G(N) glycoprotein","volume":"76","author":"Gerrard","year":"2002","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bar, S., Rommelaere, J., and Nuesch, J.P. (2013). Vesicular transport of progeny parvovirus particles through ER and Golgi regulates maturation and cytolysis. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003605"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1111\/j.1348-0421.2003.tb03424.x","article-title":"Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system","volume":"47","author":"Ito","year":"2003","journal-title":"Microb. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2933","DOI":"10.1128\/JVI.80.6.2933-2940.2006","article-title":"Rescue of infectious Rift Valley fever virus entirely from cDNA, analysis of virus lacking the NSs gene, and expression of a foreign gene","volume":"80","author":"Ikegami","year":"2006","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Kalveram, B., Lihoradova, O., Indran, S.V., and Ikegami, T. (2011). Using reverse genetics to manipulate the NSs gene of the Rift Valley fever virus MP-12 strain to improve vaccine safety and efficacy. J. Vis. Exp.","DOI":"10.3791\/3400"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/j.virol.2012.09.031","article-title":"Rift Valley fever virus NSs inhibits host transcription independently of the degradation of dsRNA-dependent protein kinase PKR","volume":"435","author":"Kalveram","year":"2013","journal-title":"Virology"},{"key":"ref_36","unstructured":"(2008). Adobe Photoshop Elements, Adobe Systems Incorporated. version 7.0 for Windows."},{"key":"ref_37","unstructured":"(2014). GraphPad Prism, GraphPad Software, Inc.. version 6.05 for Windows."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4657","DOI":"10.1016\/j.vaccine.2005.08.034","article-title":"Immunogenicity of combination DNA vaccines for Rift Valley fever virus, tick-borne encephalitis virus, Hantaan virus, and Crimean Congo hemorrhagic fever virus","volume":"24","author":"Spik","year":"2006","journal-title":"Vaccine"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.2460\/ajvr.1997.58.10.1104","article-title":"Safety and efficacy of a mutagen-attenuated Rift Valley fever virus vaccine in cattle","volume":"58","author":"Morrill","year":"1997","journal-title":"Am. J. Vet. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1016\/j.vaccine.2012.10.118","article-title":"Safety and immunogenicity of recombinant Rift Valley fever MP-12 vaccine candidates in sheep","volume":"31","author":"Morrill","year":"2013","journal-title":"Vaccine"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4988","DOI":"10.1016\/j.vaccine.2013.08.003","article-title":"Immunogenicity of a recombinant Rift Valley fever MP-12-NSm deletion vaccine candidate in calves","volume":"31","author":"Morrill","year":"2013","journal-title":"Vaccine"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3762","DOI":"10.1128\/JVI.02483-08","article-title":"Electron cryo-microscopy and single-particle averaging of Rift Valley fever virus: Evidence for GN-GC glycoprotein heterodimers","volume":"83","author":"Huiskonen","year":"2009","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Rusu, M., Bonneau, R., Holbrook, M.R., Watowich, S.J., Birmanns, S., Wriggers, W., and Freiberg, A.N. (2012). An assembly model of Rift Valley fever virus. Front Microb., 3.","DOI":"10.3389\/fmicb.2012.00254"},{"key":"ref_44","first-page":"2012","article-title":"The unique architecture of Bunyamwera virus factories around the Golgi complex","volume":"10","author":"Fontana","year":"2008","journal-title":"Genom. Comput. Sci. Virus Res."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/6\/151\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:24:22Z","timestamp":1760210662000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/8\/6\/151"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,5,24]]},"references-count":44,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2016,6]]}},"alternative-id":["v8060151"],"URL":"https:\/\/doi.org\/10.3390\/v8060151","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,5,24]]}}}