In vitro synthesis and processing of herpes simplex virus type 2 gG-2, using cell-free transcription and translation systems.
Translation of in vitro-synthesized herpes simplex virus type 2 (HSV-2) gG-2 mRNA in a reticulocyte lysate system was used to study the processing of HSV-2 gG-2. In the presence of canine pancreatic microsomal membranes, a single species that is protected from trypsin digestion was detected. This product comigrates with the 104,000-Mr (104K) high mannose intermediate seen in HSV-2-infected-cell lysates. Endo-beta-N-acetylglucosaminidase H treatment of the in vitro-synthesized 104K protein yielded a single product migrating at 100 K. The 72K and 31K cleavage products of gG-2 were not observed in the in vitro system. These data show that the molecular weight of the nonglycosylated form of the gG-2 protein is 100,000 and that the cotranslational processing of this protein in the endoplasmic reticulum yields the 104K high-mannose intermediate.
Weldon
S K
SK
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, Shreveport 71130-3932.
Su
H K
HK
Fetherston
J D
JD
Courtney
R J
RJ
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D008970:Molecular Weight; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D012156:Reticulocytes / Q000378:metabolism; D018139:Simplexvirus / Q000235:genetics*; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*
D012333:RNA, Messenger; D014759:Viral Envelope Proteins; C045464:glycoprotein G, herpes simplex virus type 2