Related Experiment Videos
Myristylation of the envelope glycoprotein of vesicular stomatitis virus
1Department of Pediatrics, Harvard Medical School, Children's Hospital, Boston, MA 02115.
Abstract:
The envelope glycoprotein G of the Indiana serotype of vesicular stomatitis virus was modified not only by palmitylation, but also by myristylation, both occurring at the carboxy-terminal segment. When infected Chinese hamster ovary cells were grown in medium containing [3H]-myristate, the G protein and Ga2, the membrane-anchoring fragment containing about 71 amino acids, were labeled. This was shown by immunoprecipitation of cell lysates or extracellular fractions of infected cultures. Thin-layer chromatography of the fatty acid fraction released from G by hydrochloride hydrolysis showed that myristate, per se, was bound to G. The sensitivity of the fatty acid linkage to KOH/methanol indicated that [3H]-myristate was linked to the protein via ester or thioester bond(s). In contrast, the G protein of New Jersey serotype was not myristylated.
Insights
Vesicular stomatitis virus G protein undergoes myristylation, a fatty acid modification. This modification was observed in the Indiana serotype but not the New Jersey serotype.
Area of Science:
- Virology
- Molecular Biology
- Protein Biochemistry
Background:
- The envelope glycoprotein G is crucial for vesicular stomatitis virus (VSV) infection.
- Lipid modifications, such as palmitylation, are known to affect viral protein function.
- The specific lipid modifications of VSV G protein require further investigation.
Purpose of the Study:
- To investigate the lipid modifications of the Indiana serotype vesicular stomatitis virus envelope glycoprotein G.
- To determine if myristylation occurs on the G protein and identify the site of modification.
- To compare the lipid modification profile of the Indiana serotype with the New Jersey serotype.
Main Methods:
- Chinese hamster ovary cells infected with VSV (Indiana serotype) were cultured in the presence of [3H]-myristate.
- Immunoprecipitation was used to isolate the G protein and its membrane-anchoring fragment (Ga2) from cell lysates and extracellular fractions.
- Thin-layer chromatography and acid hydrolysis were employed to analyze the fatty acid linkage to the G protein.
Main Results:
- The G protein and the Ga2 fragment of the Indiana serotype VSV were successfully labeled with [3H]-myristate.
- Analysis confirmed that myristate was directly bound to the G protein.
- The fatty acid linkage was sensitive to KOH/methanol, suggesting ester or thioester bonds.
- The G protein of the New Jersey serotype VSV did not exhibit myristylation.
Conclusions:
- The envelope glycoprotein G of the Indiana serotype of vesicular stomatitis virus is myristylated at its carboxy-terminal segment.
- Myristylation involves the covalent attachment of myristate via ester or thioester bonds.
- This lipid modification is specific to the Indiana serotype, as the New Jersey serotype G protein is not myristylated.