Evidence for a separate signal sequence for the carboxy-terminal envelope glycoprotein E1 of Semliki forest virus.
When Semliki Forest virus temperature-sensitive mutant ts-3 was grown at the restrictive temperature an aberrant nascent cleavage of the 130,000-dalton structural polyprotein took place relatively frequently. This cleavage yielded an abnormal 86,000-dalton fusion protein (p86) consisting of the amino-terminal capsid protein linked to the amino acid sequences of envelope protein p62 (a precursor of E3 and E2). The other cleavage product was the carboxy-terminal envelope protein E1. p86 was not glycosylated and was sensitive to the action of protease in the microsomal fraction, whereas E1 was glycosylated and protected from proteases, indicating that it had been segregated into the cysternal side of the microsomal vesicles. All attempts to show the E1 protein at the cell surface have failed so far, suggesting that it remains associated with intracellular membranes. When ts-3-infected cells labeled at the restrictive temperature were shifted to the permissive temperature the only labeled protein released with the virus particles was E1, indicating that E1, synthesized at the restrictive temperature, was competent to participate in the virus assembly. These results suggest strongly that there are two separate signal sequences for the envelope proteins of Semliki Forest virus. One follows the capsid protein as shown previously, and the other is for the carboxy-terminal E1. Even if the insertion of the amino-terminal envelope protein (p62) fails due to a cleavage defect, the other signal sequence can operate independently to guide the E1 through the endoplasmic reticulum membrane.
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D009154:Mutation; D011498:Protein Precursors / Q000378:metabolism; D012672:Semliki forest virus / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D006023:Glycoproteins; D008565:Membrane Proteins; D011498:Protein Precursors; D014764:Viral Proteins