title: Characterization of an RNA aptamer against HPV-16 L1 virus-like particles.
abstract: The human papillomavirus (HPV) capsid is mainly composed of the L1 protein that can self-assemble into virus-like particles (VLPs) that are structurally and immunologically similar to the infectious virions. We report here the characterization of RNA aptamers that recognize baculovirus-produced HPV-16 L1 VLPs. Interaction and slot-blot binding assays showed that all isolated aptamers efficiently bound HPV-16 VLPs, although the Sc5-c3 aptamer showed the highest specificity and affinity (Kd=0.05 pM). Sc5-c3 secondary structure consisted of a hairpin with a symmetric bubble and an unstructured 3'end. Biochemical and genetic analyses showed that the Sc5-c3 main loop is directly involved on VLPs binding. In particular, binding specificity appeared mediated by five non-consecutive nucleotide positions. Experiments using bacterial-produced HPV-16 L1 resulted in low Sc5-c3 binding, suggesting that recognition of HPV-16 L1 VLPs relies on quaternary structure features not present in bacteria-produced L1 protein. Sc5-c3 produced specific and stable binding to HPV-16 L1 VLPs even in biofluid protein mixes and thus it may provide a potential diagnostic tool for active HPV infection.
authors.affiliation: 1 Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del I.P.N. , México D.F., México .
authors.forename: Ana Gabriela
authors.initials: AG
authors.lastname: Leija-Montoya
authors.forename: María Luisa
authors.initials: ML
authors.lastname: Benítez-Hess
authors.forename: Julia Dolores
authors.initials: JD
authors.lastname: Toscano-Garibay
authors.forename: Luis Marat
authors.initials: LM
authors.lastname: Alvarez-Salas
mesh_terms: D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000378:metabolism*; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D036022:Capsid Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D052162:Human papillomavirus 16 / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D014771:Virion / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16
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