Association of vesicular stomatitis virus glycoprotein with virion membrane: characterization of the lipophilic tail fragment

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mentioning

confidence: 99%

Antigenicity, function, and conformation of synthetic oligopeptides corresponding to amino-terminal sequences of wild-type and mutant matrix proteins of vesicular stomatitis virus

Shipley

Pal

Wagner

1988

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“…Vesicular stomatitis virus (VSV) is an enveloped, negative-strand RNA virus with five virus-coded structural proteins (51). The N, L, and NS proteins are tightly associated with the ribonucleoprotein (RNP) core (51), whereas the G protein is an integral membrane protein that comprises the glycoprotein spikes on the surface of the viral envelope (4,48,58). The matrix (M) protein is associated with both the envelope and the RNP core structure (58); it is, however, not exposed on the exterior surface of the virion, since it is refractory to protease digestion and lactoperoxidase iodination (4,30,31,36,48).…”

mentioning

confidence: 99%

Mapping regions of the matrix protein of vesicular stomatitis virus which bind to ribonucleocapsids, liposomes, and monoclonal antibodies

Ogden¹,

Pal²,

Wagner³

1986

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The matrix (M) protein of vesicular stomatitis virus (VSV) appears to function as a bridge between the ribonucleocapsid (RNP) core and the envelope in assembly of the virion. Two such properties would necessitate at least one site for interaction with the nucleocapsid and one with the envelope. In this study M protein was found to mediate the in vitro binding to RNP cores of phospholipid vesicles, representing membrane structures. The M protein could bind initially to either the vesicles or the RNP cores to promote RNP-vesicle association. A trypsin-resistant fragment (MT) of M protein, missing the initial 43 amino acids from its amino terminus, reconstituted with acidic phospholipid vesicles with the same binding efficiency as did whole M protein, suggesting that the carboxy-terminal 81% retained those regions of the M protein which interact with a lipid bilayer. The MT protein, however, was considerably less efficient than intact M protein as an inhibitor of in vitro virus transcription; almost 2.5-fold more MT protein than intact M protein was required for 50% inhibition of VSV transcription, indicating that a site for interaction with the RNP core may have been lost. A monoclonal antibody which is able to reverse the in vitro inhibition of transcription by M protein did not react by immunoblotting with MT protein. Partial tryptic digests of the M protein probed with this monoclonal antibody indicated that epitope 1 lies between amino acid residues 18 and 43. This region appears to be a site that promotes interaction of the M protein with the RNP core of VSV. Monoclonal antibodies to epitopes 2 and 3, which exhibit some overlap in binding to M protein but do not reverse transcription inhibition, were mapped by cleavage with N-chlorosuccinimide at regions in a carboxy direction from epitope 1. * Corresponding author. t Present address: Biotherapeutics Incorporated, Franklin, TN 37064.G, and M) take independent paths to the site of virus assembly at the plasma membrane of the infected cell (1,2,6,16,22,23,34); synthesis of M protein is the rate-limiting step in this maturation process (53). Removal of M protein from the RNP core yields a relaxed, highly extended nucleocapsid structure, while condensation of the RNP into a tightly coiled skeleton is mediated through the association of M protein with the nucleocapsid (20,37,38). Appearance of M protein at the cell membrane causes decreased mobility of the already-inserted G protein (46). Protein cross-linking studies with intact virions revealed the formation of G-M and M-N but not G-N heterodimers (18). These studies suggest that M protein functions as a bridge between the cytoplasmic RNP structures and those regions of the cellular membrane into which the G protein has been inserted.The strong interaction between M protein and the RNP core enables M protein to act as an endogenous inhibitor of viral transcription. Transcription reactions with detergentdisrupted whole virions demonstrate a virus-concentrationdependent regulatory effect which was attributed to an ...

“…A host-derived lipid bilayer surrounds the nucleocapsid-matrix protein structure (6). Embedded in the bilayer are copies of a virus-coded glycoprotein (G) (36). A small number of specific host glycoproteins are also found in the viral membrane (16,27).…”

mentioning

confidence: 99%

Assembly of vesicular stomatitis virus: distribution of the glycoprotein on the surface of infected cells

Jacobs¹,

Penhoet²

1982

This study demonstrates that the glycoprotein of vesicular stomatitis virus clusters in the plasma membrane of infected Chinese hamster lung cells during morphogenesis and suggests that viral nucleocapsids are required for this clustering. A mutant virus (ts E-1) which is temperature sensitive for the synthesis of viral nucleocapsids but not viral membrane proteins was used. The surface distribution of the viral glycoprotein in cells infected by this virus was determined by a specific indirect immunoferritin stain. Early in infection at permissive temperatures, the glycoprotein was randomly distributed on membrane ghosts. Later, clusters of ferritin the size and shape of virus particles were seen. In contrast, ghosts prepared from virus-infected cells maintained at a restrictive temperature always had a random distribution of viral glycoprotein.Integral membrane proteins span the membranes with which they are associated and may participate in significant molecular interactions on both sides of the membrane. In many cases, these proteins are freely diffusible in the plane of the membrane (37). An increasing number of membrane proteins, however, are found to be distributed over the surface of the cell in a nonrandom fashion. For example, Ash et al. (1) have shown that several antibody-patched integral membrane proteins are arranged in linear arrays on the surface of cultured fibroblasts, specific receptor molecules are frequently localized in defined regions of the membrane, and viral glycoproteins of lipid-containing viruses are localized in regions of the membrane active in virus assembly (8). The molecular interactions which lead to the nonrandom distribution of integral proteins are currently under active study in a number of laboratories. Evidence is accumulating which indicates that the linear arrays of membrane proteins visible on the surface of fibroblasts result from interactions of domains of these proteins present on the cytoplasmic surface with elements of the cytoskeleton (1, 14,22). In the case of the best-studied receptor system, that of the low-density lipoproteins, the molecules appear to be inserted into the membrane at random sites and then to migrate laterally in the plane of the membrane until they reach a "coated pit." A domain of the receptor presumed to be on the cytoplasmic side t Present address: