A comparison of genomic and viral expressed sequences of a virus gene manipulation TLR responses
Year: 2014
Type: dissertation
Abstract: The I329L ORF of the African swine fever virus is a host evasion gene coding for a transmembrane protein inhibiting the induction of interferon through activation of Toll Like Receptor 3 (TLR3). Additionally, by sharing sequence similarities with TLR3, and also inhibiting TRIF mediated NF-kb activation, the protein has been characterized as a viral TLR3 antagonist. The key observation formed backing the project was that the I329L protein expressed in transfected cells not only yielded the predicted 50 kDa component, but also another one of 35 kDa. Thus the objective of this work was to explain the origin of the 35 kDa component. Two hypotheses were proposed to explain the origin of the 35 kDa molecule: (a) Translation of the I329L from a second AUG, predicted to yield a molecule of the expected size. (b) Proteolytic degradation of the 50 kDa full length protein, as indeed has been observed for other TLRs. Clear cut evidence for the first hypothesis was obtained as an I329L mutated at the second AUG only yielded a 50 kDa component in transfected cells. Importantly, both full length and second AUG 35 kDa constructs induced a similar inhibition of TLR3 signalling in luciferase reporter assays. Moreover, the full length and 35 kDa components were found to similarly localize to the endosome after activation of the TLR3 with double stranded RNA.
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2274528702