Role of viral envelope sialic acid in membrane fusion mediated by the vesicular stomatitis virus envelope glycoprotein.
Fusion of vesicular stomatitis virus (VSV) with cells and liposomes before and after treatment with neuraminidase was studied using the R18 dequenching assay. Desialylation of VSV significantly enhanced the extent of fusion with Vero cells but affected neither the pH dependence nor the binding of VSV to Vero cells. The enhanced fusion of asialo-VSV was observed both at the plasma membrane as well as via the endocytic pathway. Both VSV and asialo-VSV fused with liposomes made of neutral phospholipid, but only asialo-VSV fused with liposomes containing a 1:1 mixture of neutral and negatively charged phospholipid. To examine factors which contribute to the extent of fusion, we analyzed the various activation and inactivation reactions that take place as a result of low-pH triggering of VSV prebound to the target membrane. Lag times for the onset of fusion were similar for VSV and asialo-VSV, indicating that desialylation did not affect the activation reactions. However, exposure of VSV bound to target membranes at pH 6.5 for 400 s led to considerable inactivation, whereas little inactivation was seen after desialylation of VSV. These results are analyzed in terms of a model which allows us to determine which components of the overall fusion process are dominated by viral envelope sialic acid.
Puri
A
A
Section on Membrane Structure and Function, LMMB, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Grimaldi
S
S
Blumenthal
R
R
D000818:Animals; D002462:Cell Membrane / Q000382:microbiology / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008561:Membrane Fusion / Q000187:drug effects / Q000502:physiology*; D008562:Membrane Glycoproteins; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000494:pharmacology; D012794:Sialic Acids / Q000502:physiology*; D014709:Vero Cells; D014721:Vesicular stomatitis Indiana virus / Q000502:physiology; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000502:physiology*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D012794:Sialic Acids; D014759:Viral Envelope Proteins; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid