Effect of Certain Polypeptides on the Biological Activities of Measles Virus

Norrby, Erling; Sievertsson, Hans

doi:10.1128/aac.5.4.426

Cited by 10 publications

(4 citation statements)

References 11 publications

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“…The peptide also blocked hemolysis of African green monkey red blood cells mediated by this strain of MeV. Similar results were also demonstrated with Z-D-Phe-L-Phe-L-(NO 2 )Arg (49,50). The carbobenzoxy (Z) and D-Phe moieties are critical to the activity of FIP, and Z-L-Phe-L-Phe-Gly was shown to be a less potent inhibitor (46).…”

Section: Resultssupporting

confidence: 68%

“…The peptide also blocked hemolysis of African green monkey red blood cells mediated by MeV Edmonston-derived glycoproteins. Similar results had previously been demonstrated with Z-D-Phe-L-Phe-L(NO 2 )Arg (47)(48)(49)(50). Past experiments were performed with the Edmonston strain of the virus, which uses a CD46 receptor that is expressed on most primate cells and monkey erythrocytes (51,52).…”

supporting

confidence: 74%

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Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy- d -Phe- l -Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide

Delpeut

Noyce

et al. 2017

J Virol

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The inhibitors carbobenzoxy (Z)-d-Phe-l-Phe-Gly (fusion inhibitor peptide [FIP]) and 4-nitro-2-phenylacetyl amino-benzamide (AS-48) have similar efficacies in blocking membrane fusion and syncytium formation mediated by measles virus (MeV). Other homologues, such as Z-d-Phe, are less effective but may act through the same mechanism. In an attempt to map the site of action of these inhibitors, we generated mutant viruses that were resistant to the inhibitory effects of Z-d-Phe-l-Phe-Gly. These 10 mutations were localized to the heptad repeat B (HRB) region of the fusion protein, and no changes were observed in the viral hemagglutinin, which is the receptor attachment protein. Mutations were validated in a luciferase-based membrane fusion assay, using transfected fusion and hemagglutinin expression plasmids or with syncytium-based assays in Vero, Vero-SLAM, and Vero-Nectin 4 cell lines. The changes I452T, D458N, D458G/V459A, N462K, N462H, G464E, and I483R conferred resistance to both FIP and AS-48 without compromising membrane fusion. The inhibitors did not block hemagglutinin protein-mediated binding to the target cell. Edmonston vaccine/laboratory and IC323 wild-type strains were equally affected by the inhibitors. Escape mutations were mapped upon a three-dimensional (3D) structure modeled from the published crystal structure of parainfluenzavirus 5 fusion protein. The most effective mutations were situated in a region located near the base of the globular head and its junction with the alpha-helical stalk of the prefusion protein. We hypothesize that the fusion inhibitors could interfere with the structural changes that occur between the prefusion and postfusion conformations of the fusion protein. Due to lapses in vaccination worldwide that have caused localized outbreaks, measles virus (MeV) has regained importance as a pathogen. Antiviral agents against measles virus are not commercially available but could be useful in conjunction with MeV eradication vaccine programs and as a safeguard in oncolytic viral therapy. Three decades ago, the small hydrophobic peptide Z-d-Phe-l-Phe-Gly (FIP) was shown to block MeV infections and syncytium formation in monkey kidney cell lines. The exact mechanism of its action has yet to be determined, but it does appear to have properties similar to those of another chemical inhibitor, AS-48, which appears to interfere with the conformational change in the viral F protein that is required to elicit membrane fusion. Escape mutations were used to map the site of action for FIP. Knowledge gained from these studies could help in the design of new inhibitors against morbilliviruses and provide additional knowledge concerning the mechanism of virus-mediated membrane fusion.

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Section: Resultssupporting

confidence: 68%

supporting

confidence: 74%

Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy- d -Phe- l -Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide

Delpeut

Noyce

et al. 2017

J Virol

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“…4E). After infection, the inoculum was removed and replaced with fresh medium containing 200 M fusion-inhibitory peptide (FIP) (Z-fFG; carbobenzoxyl-D-phenylalanine-L-phenylalanine-glycine) to prevent secondary infection and syncytium formation in cells (22,78,79). Twenty hours later, numbers of GFP-positive MeV-infected cells were quantified by FACS analysis (Fig.…”

Section: Generation and Analysis Of Recombinant Wtmev Expressing The mentioning

confidence: 99%

Measles Virus Enters Breast and Colon Cancer Cell Lines through a PVRL4-Mediated Macropinocytosis Pathway

Delpeut

Sisson

Black

et al. 2017

J Virol

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Measles virus (MeV) is a member of the family that causes a highly contagious respiratory disease but has emerged as a promising oncolytic platform. Previous studies of MeV entry focused on the identification of cellular receptors. However, the endocytic and trafficking pathways utilized during MeV entry remain poorly described. The contribution of each endocytic pathway has been examined in cells that express the MeV receptors SLAM (signaling lymphocyte-activating molecule) and PVRL4 (poliovirus receptor-like 4) (nectin-4). Recombinant MeVs expressing either firefly luciferase or green fluorescent protein together with a variety of inhibitors were used. The results showed that MeV uptake was dynamin independent in the Vero.hPVRL4, Vero.hSLAM, and PVRL4-positive MCF7 breast cancer cell lines. However, MeV infection was blocked by 5-(-ethyl--propyl)amiloride (EIPA), the hallmark inhibitor of macropinocytosis, as well as inhibitors of actin polymerization. By using phalloidin staining, MeV entry was shown to induce actin rearrangements and the formation of membrane ruffles accompanied by transient elevated fluid uptake. Small interfering RNA (siRNA) knockdown of p21-activated kinase 1 (PAK1) demonstrated that MeV enters both Vero.hPVRL4 and Vero.hSLAM cells in a PAK1-independent manner using a macropinocytosis-like pathway. In contrast, MeV entry into MCF7 human breast cancer cells relied upon Rac1 and its effector PAK1 through a PVRL4-mediated macropinocytosis pathway. MeV entry into DLD-1 colon and HTB-20 breast cancer cells also appeared to use the same pathway. Overall, these findings provide new insight into the life cycle of MeV, which could lead to therapies that block virus entry or methods that improve the uptake of MeV by cancer cells during oncolytic therapy. In the past decades, measles virus (MeV) has emerged as a promising oncolytic platform. Previous studies concerning MeV entry focused mainly on the identification of putative receptors for MeV. Nectin-4 (PVRL4) was recently identified as the epithelial cell receptor for MeV. However, the specific endocytic and trafficking pathways utilized during MeV infections are poorly documented. In this study, we demonstrated that MeV enters host cells via a dynamin-independent and actin-dependent endocytic pathway. Moreover, we show that MeV gains entry into MCF7, DLD-1, and HTB-20 cancer cells through a PVRL4-mediated macropinocytosis pathway and identified the typical cellular GTPase and kinase involved. Our findings provide new insight into the life cycle of MeV, which may lead to the development of therapies that block the entry of the virus into the host cell or alternatively promote the uptake of oncolytic MeV into cancer cells.

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“…Subsequently, Norrby and Sievertsson showed that the oligopeptide [Z-D-Phe-L-Phe-L-(N0 2)Arg] inhibits virus-induced cell fusion and hemolysis as well as virus infectivity at the level of penetration (155,156). The amino acids in this tripeptide inhibitor resembled those of the N-terminal region of the Fl polypeptides of paramyxoviruses, and this raised the possibility that this similarity is responsible for the inhibition of the fusion reaction by this compound.…”

Section: Fl Hvjmentioning

confidence: 99%

Assembly of Paramyxoviruses

Matsumoto

1982

Microbiology and Immunology

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Effect of Certain Polypeptides on the Biological Activities of Measles Virus

Cited by 10 publications

References 11 publications

Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy- d -Phe- l -Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide

Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy- d -Phe- l -Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide

Measles Virus Enters Breast and Colon Cancer Cell Lines through a PVRL4-Mediated Macropinocytosis Pathway

Assembly of Paramyxoviruses

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