Biomaterial Database

Individual roles of N-linked oligosaccharide chains in intracellular transport of the paramyxovirus SV5 fusion protein. 1995

S Bagai, and R A Lamb

Howard Hughes Medical Institute, Northwestern University, Evanston, Illinois 60208-3500, USA.

The role of N-linked glycosylation in the assembly, intracellular transport, and fusion activity of the paramyxovirus SV5 fusion (F) protein was examined. Each of the six potential glycosylation sites in the F protein was individually removed by oligonucleotide-directed mutagenesis on a cDNA clone encoding the SV5 F protein. When the mutant F proteins were expressed in eukaryotic cells using the vaccinia virus-T7 transient expression system they all had a significant change in gel mobility, indicating that all six sites in the F protein are used for the addition of N-linked oligosaccharides. All of the mutant F proteins could form a homooligomer. Removal of individual carbohydrate chains from the F2 subunit had little effect on the surface expression of the F protein. However, removal of individual carbohydrate chains from the F1 subunit had deleterious effects, which ranged from a partial delay in intracellular transport and decreased stability of the protein to severe transport delays and acute instability of the F protein.

UI	MeSH Term	Description	Entries
D009844	Oligosaccharides	Carbohydrates consisting of between two (DISACCHARIDES) and ten MONOSACCHARIDES connected by either an alpha- or beta-glycosidic link. They are found throughout nature in both the free and bound form.	Oligosaccharide
D004279	DNA, Viral	Deoxyribonucleic acid that makes up the genetic material of viruses.	Viral DNA
D006031	Glycosylation	The synthetic chemistry reaction or enzymatic reaction of adding carbohydrate or glycosyl groups. GLYCOSYLTRANSFERASES carry out the enzymatic glycosylation reactions. The spontaneous, non-enzymatic attachment of reducing sugars to free amino groups in proteins, lipids, or nucleic acids is called GLYCATION (see MAILLARD REACTION).	Protein Glycosylation,Glycosylation, Protein
D006056	Golgi Apparatus	A stack of flattened vesicles that functions in posttranslational processing and sorting of proteins, receiving them from the rough ENDOPLASMIC RETICULUM and directing them to secretory vesicles, LYSOSOMES, or the CELL MEMBRANE. The movement of proteins takes place by transfer vesicles that bud off from the rough endoplasmic reticulum or Golgi apparatus and fuse with the Golgi, lysosomes or cell membrane. (From Glick, Glossary of Biochemistry and Molecular Biology, 1990)	Golgi Complex,Apparatus, Golgi,Complex, Golgi
D006367	HeLa Cells	The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays.	Cell, HeLa,Cells, HeLa,HeLa Cell
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001693	Biological Transport, Active	The movement of materials across cell membranes and epithelial layers against an electrochemical gradient, requiring the expenditure of metabolic energy.	Active Transport,Uphill Transport,Active Biological Transport,Biologic Transport, Active,Transport, Active Biological,Active Biologic Transport,Transport, Active,Transport, Active Biologic,Transport, Uphill
D014760	Viral Fusion Proteins	Proteins, usually glycoproteins, found in the viral envelopes of a variety of viruses. They promote cell membrane fusion and thereby may function in the uptake of the virus by cells.	Fusion Proteins, Viral,Viral Fusion Glycoproteins,F Protein (Sendai Virus),F Protein Measles Virus,F Protein Newcastle Disease Virus,F Protein SV,F-Glycoprotein SV,F1 Polypeptide (Paramyxovirus),Fusion Glycoprotein, Viral,Fusion VP1 Protein,Glycoprotein, Viral Fusion,Measles Fusion Protein,Mumps Virus Fusion Protein,Paramyxovirus Fusion Protein,Sendai Virus Fusion Protein,Viral Fusion-GP,Virus Fusion Proteins,Fusion Glycoproteins, Viral,Fusion Protein, Measles,Fusion Protein, Paramyxovirus,Fusion Proteins, Virus,Fusion-GP, Viral,Glycoproteins, Viral Fusion,Proteins, Virus Fusion,VP1 Protein, Fusion,Viral Fusion GP,Viral Fusion Glycoprotein
D016297	Mutagenesis, Site-Directed	Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.	Mutagenesis, Oligonucleotide-Directed,Mutagenesis, Site-Specific,Oligonucleotide-Directed Mutagenesis,Site-Directed Mutagenesis,Site-Specific Mutagenesis,Mutageneses, Oligonucleotide-Directed,Mutageneses, Site-Directed,Mutageneses, Site-Specific,Mutagenesis, Oligonucleotide Directed,Mutagenesis, Site Directed,Mutagenesis, Site Specific,Oligonucleotide Directed Mutagenesis,Oligonucleotide-Directed Mutageneses,Site Directed Mutagenesis,Site Specific Mutagenesis,Site-Directed Mutageneses,Site-Specific Mutageneses
D018076	DNA, Complementary	Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.	Complementary DNA,cDNA,cDNA Probes,Probes, cDNA