Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41

Autor(es)

Siqueira-Silva, Joselma ^[1] ; Fenel, Daphna ^[2] ; Gout, Evelyne ^[2] ; Yeda, Fernanda P. ^[1] ; Marinheiro, Juliana C. ^[1] ; Barrella, Karina M. ^[3] ; Silva, Misael L. ^[1] ; Schoehn, Guy ^{[4, 2]} ; Harsi, Charlotte M. ^[1] ; Fender, Pascal ^[4]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Ciencias, Lab Biol Mol Adenovirus, Sao Paulo - Brazil

^[2] CNRS UJF CEA UMR5075, Inst Biol Struct, Grenoble - France

^[3] Texas A&M Univ, Texas Agrilife Res Ctr, El Paso, TX - USA

^[4] CNRS UJF EMBL UMI3265, Unit Virus Host Cell Interact, FR-38042 Grenoble - France

• Tipo de documento: Artigo Científico
• Fonte: Intervirology; v. 55, n. 5, p. 349-355, 2012.
• Citações Web of Science: 0

Resumo

Objective: In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have constructed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 penton base. Methods: Recombinant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 penton base were cloned and amplified in Sf9 insect cells. Recombinant dodecahedra were expressed by coinfection of High Five (TM) cells with both baculoviruses, 72 h post-infection. Cell lysate was centrifuged on sucrose density gradient and the purified recombinant dodecahedra were recovered. Results: Analysis by negative staining electron microscopy demonstrated that chimeric dodecahedra made of the HAdV-3 penton base and decorated with the HAdV-41 SF were successfully generated. Next, recombinant dodecahedra were digested with pepsin and analyzed by Western blot. A `site-specific' proteolysis of the HAdV-41 SF was observed, while the HAdV-3 penton base core was completely digested. Conclusion: These results show that, in vitro, the HAdV-41 SF likely undergoes proteolysis in the gastrointestinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results obtained in the present study may be the basis for the development of gene therapy vectors towards the intestinal epithelium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification. Copyright (C) 2011 S. Karger AG, Basel (AU)

• Processo FAPESP: 02/13859-5 - Estudo de interação do adenovirus sorotipo 41 (HADV-41) com a célula hospedeira
• Beneficiário: Joselma Siqueira Silva
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto