Intracellular processing of the paramyxovirus F protein: critical role of the predicted amphipathic alpha helix adjacent to the fusion domain

Wang, Can; Raghu, Ganapathirama; Morrison, Trudy G.; Peeples, Mark E.

doi:10.1128/jvi.66.7.4161-4169.1992

Cited by 28 publications

(11 citation statements)

References 51 publications

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“…Interestingly, two mutations previously reported in this region are in the g and c positions on the helix (Fig. 1C, boxed letters) and result in proteins that -97 kd are temperature sensitive in folding (27).…”

Section: L155mentioning

confidence: 76%

“…2C, lane 6). with the sequence of the cytoplasmic tail of the fusion protein (27). Preliminary results with this antibody showed that, while reactivity of this antibody to wild-type protein on Western blots was low if the protein was boiled prior to electrophoresis (data not shown), proteins incubated at room temperature or heated to 50°C were readily detected.…”

Section: Iknaadvmentioning

confidence: 96%

“…Soli( changes that result in proteins that are transported to while open letters indicate changes that result in prol transported. Boxed letters indicate changes reportec (27). reeat while G128L,I151K, K155L, 1158K ( Fig.…”

mentioning

confidence: 99%

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Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion

Sergel-Germano

McQuain

Morrison

1994

J Virol

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Nonconservative mutations were introduced by site-specific mutagenesis into the fusion peptide and the adjacent heptad repeat region of the fusion protein of Newcastle disease virus in order to determine the role of both regions in the fusion activity of the protein. Mutations in both regions that allowed for proper folding and intracellular transport of the protein blocked the fusion activity of the protein when assayed in the presence of the hemagglutinin-neuraminidase protein.

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Section: L155mentioning

confidence: 76%

Section: Iknaadvmentioning

confidence: 96%

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Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion

Sergel-Germano

McQuain

Morrison

1994

J Virol

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“…To understand the mechanism of F protein-induced membrane fusion, it is important to understand the role of different domains of the F protein in mediating fusion. While it is known that the fusion peptide and adjacent heptad repeat regions have important roles in fusion Sergel-Germano et al, 1994;Wang et al, 1992), the role of the cytoplasmic tail of the F proteins is less clear. Studies with human immunodeficiency virus type 2 and simian immunodeficiency virus envelope glycoproteins (env) have shown that cytoplasmic tail truncation mutants significantly enhance syncytia formation and alter the conformation of the external domain of env (Mulligan et al, 1992;Ritter et al, 1993).…”

mentioning

confidence: 99%

Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion.

Bagai

Lamb

1996

The Journal of Cell Biology

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The role of the simian virus 5 (SV5) fusion (F) protein 20 residue COOH- terminal region, thought to represent the cytoplasmic tail, in fusion activity was examined by constructing a series of COOH-terminal truncation mutants. When the altered F proteins were expressed in eukaryotic cells, by using the vaccinia virus-T7 transient expression system, all the F proteins exhibited similar intracellular transport properties and all were expressed abundantly on the cell surface. Quantitative and qualitative cell fusion assays indicated that all of the F protein COOH-terminal truncation mutants mediated lipid mixing with similar kinetics and efficiency as that of wild-type F protein. However, the cytoplasmic content mixing activity decreased in parallel with the extent of the deletion in the F protein COOH-terminal truncation mutants. These data indicate that it is possible to separate the presumptive early step in the fusion reaction, hemifusion, and the final stage of fusion, content mixing, and that the presence of the F protein COOH-terminal region is important for the final steps of fusion.

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“…The antibody used for immunoprecipitation of the fusion protein was anti-Ftail (27). This antibody is a polyclonal rabbit antibody raised against a synthetic peptide with the sequence of the cytoplasmic tail of the fusion protein as described by Wang et al (28) .01 M NaCl) containing 1% Triton X-100, 0.5% sodium deoxycholate, and 2 mg of iodoacetamide per ml as previously described (25)(26)(27). Nuclei were removed by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

Effect of Cleavage Mutants on Syncytium Formation Directed by the Wild-Type Fusion Protein of Newcastle Disease Virus

Sergel

Razvi

et al. 1998

J Virol

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The effects of Newcastle disease virus (NDV) fusion (F) glycoprotein cleavage mutants on the cleavage and syncytium-forming activity of the wild-type F protein were examined. F protein cleavage mutants were made by altering amino acids in the furin recognition region (amino acids 112 to 116) in the F protein of a virulent strain of NDV. Four mutants were made: Q114P replaced the glutamine residue with proline; K115G replaced lysine with glycine; double mutant K115G, R113G replaced both a lysine and an arginine with glycine residues; and a triple mutant, R112G, K115G, F117L, replaced three amino acids to mimic the sequence found in avirulent strains of NDV. All mutants except Q114P were cleavage negative and fusion negative. However, addition of exogenous trypsin cleaved all mutant F proteins and activated fusion. As expected for an oligomeric protein, the fusion-negative mutants had a dominant negative phenotype: cotransfection of wild-type and mutant F protein cDNAs resulted in an inhibition of syncytium formation. The presence of the mutant F protein did not inhibit cleavage of the wild-type protein. Furthermore, evidence is presented that suggests that the mutant protein and the wild-type protein formed heterooligomers. By measuring the syncytium-forming activity of the wild-type protein at various ratios of expression of mutant and wild-type protein, results were obtained that are most consistent with the notion that the size of the functionally active NDV F protein in these assays is a single oligomer, likely a trimer. That a larger oligomer, containing a mix of both wild-type and mutant F proteins, has partial activity cannot, however, be ruled out.

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Intracellular processing of the paramyxovirus F protein: critical role of the predicted amphipathic alpha helix adjacent to the fusion domain

Cited by 28 publications

References 51 publications

Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion

Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion

Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion.

Effect of Cleavage Mutants on Syncytium Formation Directed by the Wild-Type Fusion Protein of Newcastle Disease Virus

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