title: The membrane-associated and secreted forms of the respiratory syncytial virus attachment glycoprotein G are synthesized from alternative initiation codons.
abstract: Respiratory syncytial (RS) virus synthesizes two mature forms of its attachment glycoprotein G: an anchored type II integral membrane form and a smaller form that is secreted into the medium. Here we demonstrate that these two forms are synthesized as distinct primary translation products of a single species of G protein mRNA by initiation at either of two different AUGs. Mutant cDNAs which eliminated one of the other of the two AUG codons near the 5' end of the G gene open reading frame were constructed. Analysis of the proteins synthesized from these cDNAs, either by translation of transcripts in a cell-free system or in cells infected with recombinant vaccinia viruses containing either one of the mutant cDNAs, showed that elimination of either the first or the second of these AUG codons abrogated the synthesis of the membrane-anchored or the secreted form of the protein, respectively. Additionally, two unglycosylated forms of G protein which comigrated with the unglycosylated G proteins expressed by these recombinant viruses were detected in RS virus-infected cells. Since the second AUG encodes a methionine residue that lies near the middle of the signal/anchor domain, initiation at this codon resulted in a protein with a hydrophobic amino terminus. This form of the glycoprotein was efficiently secreted from cells infected with the vaccinia virus recombinant, and the amino-terminal sequence of this protein was identical to that of G protein secreted from RS virus-infected cells. Our results demonstrate that the secreted form of RS virus G protein is produced by initiation at the second AUG codon of the G open reading frame, followed by proteolytic removal of the signal/anchor domain.
authors.affiliation: Department of Microbiology, University of Alabama Medical School, Birmingham 35294.
authors.forename: S R
authors.initials: SR
authors.lastname: Roberts
authors.forename: D
authors.initials: D
authors.lastname: Lichtenstein
authors.forename: L A
authors.initials: LA
authors.lastname: Ball
authors.forename: G W
authors.initials: GW
authors.lastname: Wertz
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D002478:Cells, Cultured; D003062:Codon; D004279:DNA, Viral; D006031:Glycosylation; D015707:HN Protein; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012136:Respiratory Syncytial Viruses / Q000378:metabolism*; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D003062:Codon; D004279:DNA, Viral; D015707:HN Protein; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G