CHARACTERIZATION OF THE GLYCOSYLATION AND SIGNAL PEPTIDE SEQUENCES WITHIN THE VACCINIA VIRUS M2 PROTEIN AND INFLUENCE ON ERK1/2 AND NF-κB ACTIVATION BY

Activation of the NF-κB transcription factor during cellular infection establishes an anti-viral state via upregulation of proinflammatory proteins. Wild-type vaccinia virus (VV) strains subvert the immune response by expressing proteins that interfere with nuclear translocation of NF-κB. In contrast, infection with an attenuated strain, MVA, results in host cell NF-κB activation. Inclusion of the M2 gene of VV into the attenuated MVA virus was shown to reduce NF-κB activation, and also ERK1/2 activation, an early event in MVA infection. Understanding the molecular basis for M2- mediated inhibition of NF-κB and ERK1/2 activation was the primary goal of this dissertation. Characterization of the M2 protein began with an examination of its expression profile during infection. Much like other immunomodulators, M2 is expressed early during infection. Analysis of the M2 amino acid sequence revealed an N-terminal signal peptide sequence, 4 putative glycosylation residues, and endoplasmic reticulum (ER) retention/retrieval sequences, indicative of an ER-localized protein