Per citar aquest document: http://ddd.uab.cat/record/112575
Altering the Ad5 packaging domain affects the maturation of the ad particles
Alba Fernández, Raúl (Universitat Autònoma de Barcelona. Centre de Biotecnologia Animal i Teràpia Gènica (CBATEG))
Cots, Dan (Universitat Autònoma de Barcelona. Centre de Biotecnologia Animal i Teràpia Gènica (CBATEG))
Ostapchuk, Philomena (Stony Brook University. Department of Molecular Genetics and Microbiology (Nova York, Estats Units d'Amèrica))
Bosch i Merino, Assumpció (Universitat Autònoma de Barcelona. Centre de Biotecnologia Animal i Teràpia Gènica (CBATEG))
Hearing, Patrick (Stony Brook University. Department of Molecular Genetics and Microbiology (Nova York, Estats Units d'Amèrica))
Chillón Rodríguez, Miguel (Universitat Autònoma de Barcelona. Centre de Biotecnologia Animal i Teràpia Gènica (CBATEG))

Data: 2011
Resum: We have previously described a new family of mutant adenoviruses carrying different combinations of attB/attP sequences from bacteriophage PhiC31 flanking the Ad5 packaging domain. These novel helper viruses have a significantly delayed viral life cycle and a severe packaging impairment, regardless of the presence of PhiC31 recombinase. Their infectious viral titers are significantly lower (100–1000 fold) than those of control adenovirus at 36 hours post-infection, but allow for efficient packaging of helper-dependent adenovirus. In the present work, we have analyzed which steps of the adenovirus life cycle are altered in attB-helper adenoviruses and investigated whether these viruses can provide the necessary viral proteins in trans. The entry of attB-adenoviral genomes into the cell nucleus early at early timepoints post-infection was not impaired and viral protein expression levels were found to be similar to those of control adenovirus. However, electron microscopy and capsid protein composition analyses revealed that attB-adenoviruses remain at an intermediate state of maturation 36 hours post-infection in comparison to control adenovirus which were fully mature and infective at this time point. Therefore, an additional 20–24 hours were found to be required for the appearance of mature attB-adenovirus. Interestingly, attB-adenovirus assembly and infectivity was restored by inserting a second packaging signal close to the right-end ITR, thus discarding the possibility that the attB-adenovirus genome was retained in a nuclear compartment deleterious for virus assembly. The present study may have substantive implications for helper-dependent adenovirus technology since helper attB-adenovirus allows for preferential packaging of helper-dependent adenovirus genomes.
Nota: Número d'acord de subvenció EC/FP7/222992
Nota: Número d'acord de subvenció NIH/AI041636
Nota: Número d'acord de subvenció CAV-4-MPS/ISCIII-PS09/02674
Nota: Número d'acord de subvenció Association Française contre les Myopathies/AFM-12277
Nota: Número d'acord de subvenció AGAUR/SGR-2009-1300
Nota: Número d'acord de subvenció Instituto Salud Carlos III/PI081162
Nota: Número d'acord de subvenció Generalitat/FI/2003-0367
Drets: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. Creative Commons
Llengua: Anglès.
Document: article ; publishedVersion
Matèria: DNA replication ; Virions ; Viral genome ; Bacterial genomics ; Vector-borne diseases ; Genomic signal processing
Publicat a: PLoS one, Vol. 6, Issue 5 (May 2011) , p. e19564, ISSN 1932-6203

DOI: 10.1371/journal.pone.0019564


10 p, 1.3 MB

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