title: Vaccinia virus L2 protein associates with the endoplasmic reticulum near the growing edge of crescent precursors of immature virions and stabilizes a subset of viral membrane proteins.
abstract: The initial step in poxvirus morphogenesis, the formation of crescent membranes, occurs within cytoplasmic factories. L2 is one of several vaccinia virus proteins known to be necessary for formation of crescents and the only one synthesized early in infection. Virus replication was unaffected when the L2R open reading frame was replaced by L2R containing an N-terminal epitope tag while retaining the original promoter. L2 colocalized with the endoplasmic reticulum (ER) protein calnexin throughout the cytoplasm of infected and transfected cells. Topological studies indicated that the N terminus of L2 is exposed to the cytoplasm with the hydrophobic C terminus anchored in the ER. Using immunogold labeling and electron microscopy, L2 was detected in tubular membranes outside factories and inside factories near crescents and close to the edge or rim of crescents; a similar labeling pattern was found for the ER luminal protein disulfide isomerase (PDI). The phenotype of L2 conditional lethal mutants and the localization of L2 suggest that it participates in elongation of crescents by the addition of ER membrane to the growing edge. Small amounts of L2 and PDI were detected within immature and mature virions, perhaps trapped during assembly. The repression of L2, as well as A11 and A17, two other proteins that are required for viral crescent formation, profoundly decreased the stability of a subset of viral membrane proteins including those comprising the entry-fusion complex. To avoid degradation, these unstable membrane proteins may need to directly insert into the viral membrane or be rapidly shunted there from the ER.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-3210, USA.
authors.forename: Liliana
authors.initials: L
authors.lastname: Maruri-Avidal
authors.forename: Andrea S
authors.initials: AS
authors.lastname: Weisberg
authors.forename: Bernard
authors.initials: B
authors.lastname: Moss
mesh_terms: D015153:Blotting, Western; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000821:virology*; D006367:HeLa Cells; D006801:Humans; D055550:Protein Stability; D014615:Vaccinia / Q000378:metabolism / Q000821:virology*; D014616:Vaccinia virus / Q000472:pathogenicity*; D014763:Viral Matrix Proteins / Q000737:chemistry* / Q000378:metabolism*; D014771:Virion / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D014763:Viral Matrix Proteins