Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Original Paper Intervirology 330631 DOI: 10.1159/000330631 Generation and Biological Properties of a Recombinant Dodecahedron Containing the Short Fiber Protein of the Human Adenovirus 41 Joselma Siqueira-Silva a Daphna Fenel b Evelyne Gout b Fernanda P. Yeda a Juliana C. Marinheiro a Karina M. Barrella c Misael L. Silva a Guy Schoehn b, d Charlotte M. Harsi a Pascal Fender d a Laboratório de Biologia Molecular de Adenovírus, Instituto de Ciências, Universidade de São Paulo, São Paulo, Brasil; b CNRS/UJF/CEA (UMR5075), Institut de Biologie Structurale, Grenoble, France; c Texas Agrilife Research Center, Texas A&M University, El Paso, Tex., USA; d CNRS/UJF/EMBL (UMI3265), Unit of Virus Host Cell Interactions, Grenoble, France blot. A ‘site-specific’ proteolysis of the HAdV-41 SF was ob- served, 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 gastrointes- tinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results ob- tained in the present study may be the basis for the develop- ment of gene therapy vectors towards the intestinal epithe- lium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification. Copyright © 2011 S. Karger AG, Basel Introduction Human adenoviruses are divided into six species (A– F), according to biological and molecular properties. The characteristics of each species determine the cell tropism and pathogenesis, resulting in respiratory, enteric or uri- nary tract infections [1, 2]. All adenoviruses are non-en- veloped, double-stranded DNA viruses, 70–90 nm in di- Key Words Human adenovirus F  Short fiber protein  Virus-like particle  Proteolysis Abstract Objective: In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have construct- ed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 penton base. Methods: Recom- binant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 penton base were cloned and amplified in Sf 9 insect cells. Recombinant dodecahedra were expressed by coinfec- tion of High Five TM cells with both baculoviruses, 72 h post- infection. Cell lysate was centrifuged on sucrose density gra- dient and the purified recombinant dodecahedra were re- covered. 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 dodeca- hedra were digested with pepsin and analyzed by Western Received: February 23, 2011 Accepted after revision: June 27, 2011 Published online: $$$ Pascal Fender CNRS/UJF/EMBL (UMI3265), Unit of Virus Host Cell Interactions 6, rue Jules Horowitz FR–38042 Grenoble Cedex (France) Tel. +33 $$$$ $$$ $$$ , E-Mail pfender  @  embl.fr © 2011 S. Karger AG, Basel 0300–5526/11/0000–0000$38.00/0 Accessible online at: www.karger.com/int INT330631.indd 1 INT330631.indd 1 24.08.2011 10:58:51 24.08.2011 10:58:51