Increased adenovirus Type 5 mediated transgene expression due to RhoB down-regulation.
Adenovirus type 5 (Ad5) is a non-enveloped DNA virus frequently used as a gene transfer vector. Efficient Ad5 cell entry depends on the availability of its primary receptor, coxsackie and adenovirus receptor, which is responsible for attachment, and integrins, secondary receptors responsible for adenovirus internalization via clathrin-mediated endocytosis. However, efficacious adenovirus-mediated transgene expression also depends on successful trafficking of Ad5 particles to the nucleus of the target cell. It has been shown that changes occurring in tumor cells during development of resistance to anticancer drugs can be beneficial for adenovirus mediated transgene expression. In this study, using an in vitro model consisting of a parental cell line, human laryngeal carcinoma HEp2 cells, and a cisplatin-resistant clone CK2, we investigated the cause of increased Ad5-mediated transgene expression in CK2 as compared to HEp2 cells. We show that the primary cause of increased Ad5-mediated transgene expression in CK2 cells is not modulation of receptors on the cell surface or change in Ad5wt attachment and/or internalization, but is rather the consequence of decreased RhoB expression. We propose that RhoB plays an important role in Ad5 post-internalization events and more particularly in Ad5 intracellular trafficking. To the best of our knowledge, this is the first study showing changed Ad5 trafficking pattern between cells expressing different amount of RhoB, indicating the role of RhoB in Ad5 intracellular trafficking.
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Dragomira
D
Majhen
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Nikolina
N
Stojanović
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Dunja
D
Vukić
Centre de Biophysique Moléculaire CNRS-UPR4301 Affiliated to the Université d'Orléans, Orléans, France.
Chantal
C
Pichon
Centre de Biophysique Moléculaire CNRS-UPR4301 Affiliated to the Université d'Orléans, Orléans, France.
Chloé
C
Leduc
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Maja
M
Osmak
Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Andreja
A
Ambriović-Ristov
D000260:Adenoviruses, Human / Q000235:genetics*; D001692:Biological Transport; D002460:Cell Line; D045744:Cell Line, Tumor; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein / Q000235:genetics; D015870:Gene Expression; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016023:Integrins / Q000235:genetics; D011991:Receptors, Virus / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes; D053585:Virus Attachment; D053586:Virus Internalization; D020743:rhoB GTP-Binding Protein / Q000235:genetics* / Q000378:metabolism
D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016023:Integrins; D011991:Receptors, Virus; D020743:rhoB GTP-Binding Protein