title: Codon optimization of the human papillomavirus 11 (HPV 11) L1 gene leads to increased gene expression and formation of virus-like particles in mammalian epithelial cells.
abstract: The 505 amino acid L1 protein of the human papillomavirus type 11 (HPV 11) is the major capsid polypeptide that has been shown to self-assemble into virus-like particles (VLPs) in vivo and in vitro. While L1 is essential for viral infection, expression studies in mammalian cells have been hampered by different codon preference between the virus and its host. To optimize L1 gene expression in mammalian cells, we converted wild-type HPV 11 L1 (11 L1wt) codons to those more common in human genes. The modified HPV 11 L1 gene (11 L1h) generated protein levels that were at least 100-fold higher than those of wild-type HPV 11 L1, while no obvious differences were seen in the level of mRNA. HPV 11 L1 protein was detected in mammalian epithelial and fibroblast cells, by immunoblotting and indirect immunofluorescence (IIF) techniques. Unlike the situation in situ, IIF revealed the presence of L1 mainly at perinuclear sites. Virus-like particles assembled intranuclearly only to a low extent, as indicated by transmission electron microscopy. DNA vaccination using the HPV 11 L1h gene yielded a drastic increase in L1-specific antibody production in mice as compared to immunization with the wild-type gene.
authors.affiliation: Research Program of Infection and Cancer, German Cancer Research Center, 69120 Heidelberg, Germany. nina.mossadegh@dkfz.de
authors.forename: Nina
authors.initials: N
authors.lastname: Mossadegh
authors.forename: Lutz
authors.initials: L
authors.lastname: Gissmann
authors.forename: Martin
authors.initials: M
authors.lastname: Müller
authors.forename: Hanswalter
authors.initials: H
authors.lastname: Zentgraf
authors.forename: Angel
authors.initials: A
authors.lastname: Alonso
authors.forename: Pascal
authors.initials: P
authors.lastname: Tomakidi
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D036022:Capsid Proteins; D002460:Cell Line; D003062:Codon; D004847:Epithelial Cells / Q000821:virology; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009856:Oncogene Proteins, Viral / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D030361:Papillomavirus Infections / Q000097:blood; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D014162:Transfection; D014611:Vaccination; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D003062:Codon; C094776:L1 protein, Human papillomavirus type 11; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D019444:Vaccines, DNA; D014765:Viral Vaccines