1887

Abstract

Intracellular processing and trafficking of the baculovirus expressed cathepsin (V-CATH), which lacks canonical targeting signals, are poorly understood. The cathepsins of multiple nucleopolyhedrovirus (AcMNPV), multiple nucleopolyhedrovirus (CfMNPV) and most other alphabaculovirus group I nucleopolyhedroviruses have well-conserved N-termini containing overlapping chymotrypsin-cleavage (Y) and myristoylation (G) motifs, which are suggestive of proteolytic signal-peptide cleavage to generate proV-CATH and subsequent acylation. To determine proteolytic N-terminal processing of V-CATH, haemagglutinin epitope-coding tags were fused to the 5′ and/or 3′ ends of AcMNPV and CfMNPV . Immunoblot analysis suggested that a small N-terminal peptide is cleaved for both viruses, indicating that is expressed as a pre-proenzyme. The two viral homologues undergo similar proteolytic processing, but have different glycosylation or other post-translational modifications. An AcMNPV V-CATH–DsRED fusion protein co-localized to the endoplasmic reticulum with an HDEL motif-containing green fluorescent protein. Based on these findings, pre-proV-CATH processing and trafficking mechanisms are postulated.

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2009-04-01
2019-11-19
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vol. , part 4, pp. 995 – 1000

Methods, references and Supplementary Table S1 (primer sequences) [ Single PDF file] (133 KB)



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