Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant
- Yale Univ. School of Medicine, New Haven, CT (USA)
The authors have used transient expression of the wild-type vesicular stomatitis virus (VSV) glycoprotein (G protein) from cloned cDNA to rescue a temperature-sensitive G protein mutant of VSV in cells at the nonpermissive temperature. Using cDNAs encoding G proteins with deletions in the normal 29-amino-acid cytoplasmic domain, they determined that the presence of either the membrane-proximal 9 amino acids or the membrane-distal 12 amino acids was sufficient for rescue of the temperature-sensitive mutant. G proteins with cytoplasmic domains derived from other cellular or viral G proteins did not rescue the mutant, nor did G proteins with one or three amino acids of the normal cytoplasmic domain. Rescue correlated directly with the ability of the G proteins to be incorporated into virus particles. This was shown by analysis of radiolabeled particles separated on sucrose gradients as well as by electron microscopy of rescued virus after immunogold labeling. Quantitation of surface expression showed that all of the mutated G proteins were expressed less efficiently on the cell surface than was wild-type G protein. However, they were able to correct for differences in rescue efficiency resulting from differences in the level of surface expression by reducing wild-type G protein expression to levels equivalent to those observed for the mutated G proteins. The results provide evidence that at least a portion of the cytoplasmic domain is required for efficient assembly of the VSV G protein into virions during virus budding.
- OSTI ID:
- 7067261
- Journal Information:
- Journal of Virology; (USA), Journal Name: Journal of Virology; (USA) Vol. 63:9; ISSN JOVIA; ISSN 0022-538X
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
CELL FLOW SYSTEMS
CELL MEMBRANES
COLLOIDS
DAYS LIVING RADIOISOTOPES
DISPERSIONS
DNA
DNA HYBRIDIZATION
ELECTRON CAPTURE RADIOISOTOPES
ELECTRON MICROSCOPY
GLYCOPROTEINS
GOLD COMPOUNDS
HYBRIDIZATION
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
LIGHT NUCLEI
MEMBRANES
MICROORGANISMS
MICROSCOPY
MOLECULAR STRUCTURE
MUTANTS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOCOLLOIDS
RADIOISOTOPES
RECOMBINANT DNA
TRANSITION ELEMENT COMPOUNDS
VIRUSES