6533b86efe1ef96bd12cb538

RESEARCH PRODUCT

Development and characterization of a 293 cell line with regulatable expression of the hepatitis B virus large envelope protein

Carsten LambertReinhild Prange

subject

Gene Expression Regulation ViralHepatitis B virusEcdysoneProtein ConformationEndoplasmic reticulumLiver cellCell MembraneCellGolgi ApparatusBiologyEndoplasmic Reticulummedicine.disease_causebiology.organism_classificationMolecular biologymedicine.anatomical_structureViral Envelope ProteinsHepadnaviridaeCell cultureVirologyMembrane topologymedicineHumansSecretionPromoter Regions GeneticCell Line Transformed

description

During the life cycle of hepatitis B virus (HBV) the large L envelope protein plays a pivotal role that is related to its peculiar dual transmembrane topology. To study the complex structure and diverse functions of L under regulated conditions of production, a human 293 cell line stably expressing L under the control of the ecdysone-inducible promoter was generated. Cells demonstrated stringent dose- and time-dependent kinetics of induction with undetectable background expression in the absence of the inducer. Temporal control of L expression allowed to trace (i) its posttranslational reorientation resulting in the mixed topology; (ii) its spatial redistribution from the endoplasmic reticulum to Golgi-like structures; and (iii) its intracellular retention in a membrane-associated configuration. On regulated overproduction, L blocked the secretion of HBV small envelope polypeptides without impairing the cell secretory apparatus. Despite the continuous high-level storage of L within the 293 cell line, no cytopathic effects could be detected. This is in contrast to ground-glass hepatocytes of chronic HBV carriers and HBV transgenic mice and may imply that the intracellular storage of L is particularly damaging to the liver cell.

yearjournalcountryeditionlanguage
2004-03-31Journal of Virological Methods
https://doi.org/10.1016/j.jviromet.2004.06.018