title: Co-translational, intraribosomal cleavage of polypeptides by the foot-and-mouth disease virus 2A peptide.
abstract: During co-translational protein import into the endoplasmic reticulum ribosomes are docked onto the translocon. This prevents inappropriate exposure of nascent chains to the cytosol and, conversely, cytosolic factors from gaining access to the nascent chain. We exploited this property of co-translational translocation to examine the mechanism of polypeptide cleavage by the 2A peptide of the foot-and-mouth disease virus. We find that the scission reaction is unaffected by placing 2A into a co-translationally targeted protein. Moreover, the portion of the polypeptide C-terminal to the cleavage site remains in the cytosol unless it contains its own signal sequence. The pattern of cleavage is consistent with the proposal that the 2A-mediated cleavage reaction occurs within the ribosome itself. In addition, our data indicate that the ribosome-translocon complex detects the break in the nascent chain and prevents any downstream protein lacking a signal sequence from gaining access to the endoplasmic reticulum.
authors.affiliation: School of Biology, Centre for Biomolecular Sciences, Biomolecular Sciences Building, University of St. Andrews, North Haugh, St. Andrews KY16 9ST, United Kingdom.
authors.forename: Pablo
authors.initials: P
authors.lastname: de Felipe
authors.forename: Lorraine E
authors.initials: LE
authors.lastname: Hughes
authors.forename: Martin D
authors.initials: MD
authors.lastname: Ryan
authors.forename: Jeremy D
authors.initials: JD
authors.lastname: Brown
mesh_terms: D004721:Endoplasmic Reticulum / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006868:Hydrolysis; D008164:Luminescent Proteins / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism*; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008164:Luminescent Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C057421:virus protein 2A; D049452:Green Fluorescent Proteins