Radtke, Christina
[Author]
;
Knittler, Michael R.
[Degree supervisor];
Tautz, Norbert
[Degree supervisor];
Tews, Birke Andrea
[Degree supervisor]Universität Greifswald
Charakterisierung der pestiviralen Glykoproteine E1 und E2
The Bovine Viral Diarrhea Virus (BVDV) belongs to the genus Pestivirus within the family of Flaviviridae. BVDV causes a severe diarrhea in cattle, especially calves, called bovine viral diarrhoea (mucosal disease). BVDV belongs to the single-stranded RNA viruses and is closely related to the pathogen of classical swine fever and border disease in sheep. Like other pestiviruses, BVDV is an enveloped virus whose envelope contains three structural proteins (Erns, E1 and E2). In this work, the glycoproteins E1 and E2 were further characterized to get first information about the viral budding mechanism. For that reason the subcellular localization, the retention as well as the membrane topology of E1 and E2 were analyzed. The subcellular localization of E1 und E2 were elucidated by performing colocalization experiments with compartment marker proteins using immunofluorescence microscopy. The results showed a localization predominantly in the ER for single- and E1-E2-precurser expressed glycoproteins. Infection studies with different BVDV strains also showed a predominantly localization in the ER for E2. To characterize the requirements for intracellular retention CD72-chimeric and mutated proteins were used. In both proteins, the transmembrane domain was responsible for retention. In further experiments, different amino acids in the C-Terminus of E1 or E2 were substituted, deleted or inserted and the influence on the retention of the mutants was analyzed. Arginine (position 355 in ...