Processing of yellow fever virus polyprotein: role of cellular proteases in maturation of the structural proteins.
The yellow fever virus (YFV) cDNA segment coding for the part of the precursor polyprotein generating the structural proteins C (capsid), prM (precursor to the membrane protein M), and E (envelope) was expressed in vitro by using the T7 promoter-polymerase transcription system coupled to translation in rabbit reticulocyte lysates. A polypeptide of the expected molecular weight was observed to accumulate in the assay and was processed into proteins C, prM, and E only when dog pancreas microsomal membranes were added to the translation system. Proteins prM and E were translocated inside the endoplasmic reticulum, where prM underwent glycosylation. Regions essential for translocation of these proteins were localized to the 18- and 15-amino-acid C-terminal hydrophobic regions of proteins C and prM, respectively. Translocation of protein prM appeared to be less efficient than that of protein E. Maturation of these proteins followed different kinetics, indicating that the prM signal is probably cleaved off more slowly. A polypeptide composed of proteins C and prM, similar to the NVx polypeptide described in yellow fever virus-infected cells, was also produced in the in vitro system in the presence of membranes. No mature protein M was detected, suggesting that the cleavage of prM to M is a late processing event mediated by a protease different from endoplasmic reticulum signalases.
Ruiz-Linares
A
A
Centre National de la Recherche Scientifique, UA 545, Institut Pasteur, Paris, France.
Cahour
A
A
Després
P
P
Girard
M
M
Bouloy
M
M
D001692:Biological Transport; D002213:Capsid / Q000378:metabolism; D003062:Codon; D006031:Glycosylation; D007700:Kinetics; D008861:Microsomes / Q000378:metabolism; D010447:Peptide Hydrolases / Q000502:physiology*; D011498:Protein Precursors / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000032:analysis; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins; D015005:Yellow fever virus / Q000378:metabolism*
D003062:Codon; D011498:Protein Precursors; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; D010447:Peptide Hydrolases