Mutations in a Carboxy-Terminal Region of Vesicular Stomatitis Virus Glycoprotein G That Affect Membrane Fusion Activity

Shokralla, Shahira; He, Yun; Wanas, Essam; Ghosh, Hara P.

doi:10.1006/viro.1997.8986

Cited by 35 publications

(33 citation statements)

References 69 publications

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“…(b) Region of the glycoprotein of VSV and RV involved in the control of the pH-dependent structural changes leading to fusion aligned with the equivalent region of VHSV. * Positions of mutations which affect the fusion process of VSV (Shokralla et al, 1998) ; † positions of mutations of the RV glycoprotein which affect the structural transitions of RV G (Gaudin et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

“…5 b). Mutations in this latter region have been shown to affect the fusion properties of RV and VSV G proteins (Gaudin et al, 1996 ;Shokralla et al, 1998), possibly by influencing the low-pH-induced conformational changes necessary for fusion activity. Our finding that (i) mutations at position 140 were always associated with a concomitant change at position 431 or 433 and that (ii) we could not select a revertant with a single mutation at this position, provided the first concrete argument that regions I and II are structurally linked in the native G protein.…”

Section: Discussionmentioning

confidence: 99%

“…In the present study, we further extended the characterization of the mutations involved in the attenuated phenotype and identified two distinct regions of G associated with virulence. One of these regions is located in or near the putative fusion peptide of VHSV described above ; the other is in a region which has been proposed to be involved in regulation of low-pH-induced conformational changes for both RV and VSV (Gaudin et al, 1996 ;Shokralla et al, 1998). Therefore, the fusion properties of VHSV (and of its variants) were also studied.…”

Section: Introductionmentioning

confidence: 99%

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Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Gaudin

Kinkelin

Benmansour

1999

Journal of General Virology

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To study the molecular basis of virulence of viral haemorrhagic septicaemia virus (VHSV), we used a cross-reactive neutralizing MAb to select MAb-resistant (MAR) mutants with reduced pathogenicity for fish. From sequence determination of the G gene of MAR mutants, attenuated laboratory variant and avirulent field strains, we identified two distant regions of the glycoprotein associated with virulence : region I (aa 135-161), homologous to the putative fusion peptide of both rabies virus (RV) and vesicular stomatitis virus (VSV), and region II (surrounding aa 431-433), homologous to RV and VSV domains controlling the conformational changes necessary for the fusion process to take place. Simultaneous mutations in both regions resulted in the most attenuated phenotype and we obtained genetic evidence that regions I and II may be structurally linked. As the MAR mutants had mutations in or near domains involved in fusion, the fusion properties of VHSV and its variants were analysed. This work allowed us to postulate that the fusion domain of VHSV is probably constituted of two distinct regions of the protein connected through a disulfide bridge between cysteines 110 and 152. Finally, we obtained evidence suggesting that the pH threshold for fusion is a determinant for virulence : restriction of fusion to a more acidic pH was associated with attenuation for the variant tr25 which had a shift of the threshold for maximal fusion from pH 6n30 (for the parental strain) to pH 6n00 ; conversely, two field strains which had maximal fusion at pH 6n60 were the most virulent.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Gaudin

Kinkelin

Benmansour

1999

Journal of General Virology

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“…Another region of G protein, encompassing residues 395 to 418 for VSV or containing residues 392 to 396 for rabies virus, has been identified as a segment that affects the fusogenic activity of the protein by influencing the low-pH-induced conformational changes (45,51). In addition, it has also been shown that not only the ectodomain segment but also the membrane anchoring domain is required for VSV fusion activity (52,53).…”

Section: Determination Of the Rhabdovirus Fusion Peptidementioning

confidence: 99%

Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

Poian

Carneiro

Stauffer

2005

Braz J Med Biol Res

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Enveloped viruses always gain entry into the cytoplasm by fusion of their lipid envelope with a cell membrane. Some enveloped viruses fuse directly with the host cell plasma membrane after virus binding to the cell receptor. Other enveloped viruses enter the cells by the endocytic pathway, and fusion depends on the acidification of the endosomal compartment. In both cases, virus-induced membrane fusion is triggered by conformational changes in viral envelope glycoproteins. Two different classes of viral fusion proteins have been described on the basis of their molecular architecture. Several structural data permitted the elucidation of the mechanisms of membrane fusion mediated by class I and class II fusion proteins. In this article, we review a number of results obtained by our laboratory and by others that suggest that the mechanisms involved in rhabdovirus fusion are different from those used by the two well-studied classes of viral glycoproteins. We focus our discussion on the electrostatic nature of virus binding and interaction with membranes, especially through phosphatidylserine, and on the reversibility of the conformational changes of the rhabdovirus glycoprotein involved in fusion. Taken together, these data suggest the existence of a third class of fusion proteins and support the idea that new insights should emerge from studies of membrane fusion mediated by the G protein of rhabdoviruses. In particular, the elucidation of the three-dimensional structure of the G protein or even of the fusion peptide at different pH's might provide valuable information for understanding the fusion mechanism of this new class of fusion proteins. Correspondence

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“…However, other studies demonstrated that amino acids 395-418 (the membrane-proximal region) have an significant influence on fusion (25), and additional studies identified region 145-164, termed the p2-like peptide, as being a pivotal domain in facilitating glycoprotein G-mediated membrane fusion (26,27). A synthetic p2 peptide was shown to mediate liposome-liposome fusion in a low pHand phosphatidylserine-dependent manner (27).…”

mentioning

confidence: 99%

Molecular Architecture of the Bipartite Fusion Loops of Vesicular Stomatitis Virus Glycoprotein G, a Class III Viral Fusion Protein

Sun

Belouzard

Whittaker

2008

Journal of Biological Chemistry

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The glycoprotein of vesicular stomatitis virus (VSV G) mediates fusion of the viral envelope with the host cell, with the conformational changes that mediate VSV G fusion activation occurring in a reversible, low pH-dependent manner. Based on its novel structure, VSV G has been classified as class III viral fusion protein, having a predicted bipartite fusion domain comprising residues Trp-72, Tyr-73, Tyr-116, and Ala-117 that interacts with the host cell membrane to initiate the fusion reaction. Here, we carried out a systematic mutagenesis study of the predicted VSV G fusion loops, to investigate the functional role of the fusion domain. Using assays of low pH-induced cell-cell fusion and infection studies of mutant VSV G incorporated into viral particles, we show a fundamental role for the bipartite fusion domain. We show that Trp-72 is a critical residue for VSV G-mediated membrane fusion. Trp-72 could only tolerate mutation to a phenylalanine residue, which allowed only limited fusion. Tyr-73 and Tyr-116 could be mutated to other aromatic residues without major effect but could not tolerate any other substitution. Ala-117 was a less critical residue, with only charged residues unable to allow fusion activation. These data represent a functional analysis of predicted bipartite fusion loops of VSV G, a founder member of the class III family of viral fusion proteins. Vesicular stomatitis virus (VSV)2 is a prototypic virus in the Rhabdoviridae, which includes many important human, animal, and plant pathogens, including Rabies virus (1). VSV is an enveloped virus that is well known to infect cells via a low pHdependent fusion reaction within endosomes (2-4). The virus contains a single envelope protein, termed the glycoprotein (G), which mediates both attachment to host cells and fusion between the virus envelope and the host cell membrane. This fusion event delivers the VSV genome into the host cell for viral replication. In addition to being a critical determinant of viral pathogenesis, VSV G has also served as an important model for protein folding and transport through the secretory pathway of cells (5). VSV G is used extensively in pseudotyped virus systems and as a delivery system for gene therapy applications (6), and, due to the fact that VSV preferentially replicates and destroys immortalized or tumorigenic cells, the virus has been of great interest as an oncolytic agent in anticancer treatment (7).The mature VSV G protein is an ϳ65-kDa type I transmembrane protein containing 511 amino acids that oligomerizes into a homotrimer during transport to the cell surface, where the trimer is then assembled into the viral particle (8). Unlike many other viral glycoproteins (9), VSV G is not subject to proteolytic priming for fusion activation. The fusogenic ability of VSV G has been of significant interest, because, unlike the proposed "spring-loaded" and essentially irreversible metastable state for the pre-fusion state of other fusion proteins such as influenza HA strain X-31 (10), VSV G is apparently fully re...

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Mutations in a Carboxy-Terminal Region of Vesicular Stomatitis Virus Glycoprotein G That Affect Membrane Fusion Activity

Cited by 35 publications

References 69 publications

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

Molecular Architecture of the Bipartite Fusion Loops of Vesicular Stomatitis Virus Glycoprotein G, a Class III Viral Fusion Protein

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