Temporal and spatial expression of the E5a protein during the differentiation-dependent life cycle of human papillomavirus type 31b.
Human papillomaviruses (HPVs) are epitheliotropic viruses, and their life cycle is intimately linked to the stratification and differentiation state of the host epithelial tissues. Defining a role for the E5 gene product in the differentiation-dependent viral life cycle has been difficult due to the lack of a suitable culture system. We used the organotypic (raft) culture system to investigate the spatial and temporal expression pattern of the E5 protein during the differentiation-dependent life cycle of HPV-31b. We report the generation of antisera specific to the HPV-31b E5a protein. The HPV-31b E5a protein was detected throughout the viral life cycle in raft cultures as determined by immunostaining analyses, and the protein was localized predominantly to the basal and granular layers. Expression of epidermal growth factor receptor or platelet-derived growth factor receptors, two proteins with which E5 has been shown to interact in cell culture, did not specifically colocalize with E5a expression. However, HPV-31b E5a expression did colocalize with the epithelial differentiation-specific marker filaggrin. The kinetics of E5a protein expression during the complete viral life cycle was analyzed by immunoblotting, and the highest level was found to be coincidental with the onset of virion morphogenesis.
Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania, 17033, USA.
T J
TJ
Mayer
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C
Meyers
D000818:Animals; D001688:Biological Products / Q000502:physiology; D004815:Epidermal Growth Factor / Q000378:metabolism; D000091344:Filaggrin Proteins; D006801:Humans; D015151:Immunoblotting; D007381:Intermediate Filament Proteins / Q000378:metabolism; D008018:Life Cycle Stages; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000378:metabolism; D014407:Tumor Cells, Cultured
D001688:Biological Products; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D009856:Oncogene Proteins, Viral; D010982:Platelet-Derived Growth Factor; C105085:viron; D004815:Epidermal Growth Factor