Nipah virus fusion protein: Influence of cleavage site mutations on the cleavability by cathepsin L, trypsin and furin

Diederich, Sandra; Dietzel, Erik; Maisner, Andrea

doi:10.1016/j.virusres.2009.07.020

Cited by 11 publications

(12 citation statements)

References 49 publications

(70 reference statements)

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“…However, the NiV proteolytic activation pathway differs from that for other systemically replicating paramyxoviruses, for example, measles virus (MV). F proteins of MV are activated by the ubiquitous Golgi apparatus protease furin during transport to the cell surface, resulting in almost complete MV F cleavage (Ͼ95%) (16,38). In contrast, the inactive precursor NiV F 0 is transported to the cell surface and must then undergo clathrin-mediated endocytosis to get in contact with its activating protease in the endolysosomal compartment.…”

Section: Niv F Cleavage Efficiency Varies Between Different Cell Typesmentioning

confidence: 99%

“…Unlike cleavage of most other paramyxovirus F proteins, NiV F processing depends on the transport of precursor NiV F 0 to the cell surface and subsequent endocytosis mediated by a tyrosine-based internalization signal in its cytoplasmic tail ( 525 YSRL) (18,77). Within the endolysosomal compartment, F 0 is then ubiquitously activated by pH-dependent proteases at its monobasic cleavage site (arginine 109) (16,48). After cleavage and release of the hydrophobic peptide at the N terminus of F 1 , the fusion-active F 1 -F 2 form is recycled to the cell surface, where it can induce syncytium formation or is incorporated into budding virus particles.…”

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confidence: 99%

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Activation of the Nipah Virus Fusion Protein in MDCK Cells Is Mediated by Cathepsin B within the Endosome-Recycling Compartment

Diederich

Sauerhering

Weis

et al. 2012

J Virol

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c Proteolytic activation of the fusion protein of the highly pathogenic Nipah virus (NiV F) is a prerequisite for the production of infectious particles and for virus spread via cell-to-cell fusion. Unlike other paramyxoviral fusion proteins, functional NiV F activation requires endocytosis and pH-dependent cleavage at a monobasic cleavage site by endosomal proteases. Using prototype Vero cells, cathepsin L was previously identified to be a cleavage enzyme. Compared to Vero cells, MDCK cells showed substantially higher F cleavage rates in both NiV-infected and NiV F-transfected cells. Surprisingly, this could not be explained either by an increased F endocytosis rate or by elevated cathepsin L activities. On the contrary, MDCK cells did not display any detectable cathepsin L activity. Though we could confirm cathepsin L to be responsible for F activation in Vero cells, inhibitor studies revealed that in MDCK cells, cathepsin B was required for F-protein cleavage and productive replication of pathogenic NiV. Supporting the idea of an efficient F cleavage in early and recycling endosomes of MDCK cells, endocytosed F proteins and cathepsin B colocalized markedly with the endosomal marker proteins early endosomal antigen 1 (EEA-1), Rab4, and Rab11, while NiV F trafficking through late endosomal compartments was not needed for F activation. In summary, this study shows for the first time that endosomal cathepsin B can play a functional role in the activation of highly pathogenic NiV.

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Section: Niv F Cleavage Efficiency Varies Between Different Cell Typesmentioning

confidence: 99%

mentioning

confidence: 99%

Activation of the Nipah Virus Fusion Protein in MDCK Cells Is Mediated by Cathepsin B within the Endosome-Recycling Compartment

Diederich

Sauerhering

Weis

et al. 2012

J Virol

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“…Some viruses use the protease activity of particular cellular enzymes localized in the endosomes or at the cellular surface for activating their EnvGP. The Ebola filovirus (for which the GP1-GP2 envelope glycoprotein is cleaved by the furine in the producer cells) (Chandran et al, 2005;Schornberg et al, 2006), the HeV (Pager and Dutch, 2005), NiV (Diederich et al, 2009), or the SARS-CoV and MHV-2 coronaviruses (for which the S spikes is not cleaved in the producer cells) (Huang et al, 2006;Qiu et al, 2006;Simmons et al, 2005) require the cysteins lysosomal proteases, the L or B cathepsine (CatL or CatB) for their entry process. After virus uptake following angiotensin-converting enzyme 2 receptor binding, cathepsin L-mediated proteolysis induces conformational changes in the SARS-CoV S glycoprotein to trigger the endosomal membrane fusion process (Simmons et al, 2005).…”

Section: Fusion Activation By Proteasesmentioning

confidence: 99%

Cell Entry of Enveloped Viruses

Cosset

Lavillette

2011

Advances in Genetics

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Enveloped viruses penetrate their cell targets following the merging of their membrane with that of the cell. This fusion process is catalyzed by one or several viral glycoproteins incorporated on the membrane of the virus. These envelope glycoproteins (EnvGP) evolved in order to combine two features. First, they acquired a domain to bind to a specific cellular protein, named "receptor." Second, they developed, with the help of cellular proteins, a function of finely controlled fusion to optimize the replication and preserve the integrity of the cell, specific to the genus of the virus. Following the activation of the EnvGP either by binding to their receptors and/or sometimes the acid pH of the endosomes, many changes of conformation permit ultimately the action of a specific hydrophobic domain, the fusion peptide, which destabilizes the cell membrane and leads to the opening of the lipidic membrane. The comprehension of these mechanisms is essential to develop medicines of the therapeutic class of entry inhibitor like enfuvirtide (Fuzeon) against human immunodeficiency virus (HIV). In this chapter, we will summarize the different envelope glycoprotein structures that viruses develop to achieve membrane fusion and the entry of the virus. We will describe the different entry pathways and cellular proteins that viruses have subverted to allow infection of the cell and the receptors that are used. Finally, we will illustrate more precisely the recent discoveries that have been made within the field of the entry process, with a focus on the use of pseudoparticles. These pseudoparticles are suitable for high-throughput screenings that help in the development of natural or artificial inhibitors as new therapeutics of the class of entry inhibitors.

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“…To test our hypothesis, we used protease inhibitors to identify cathepsin L to be the protease responsible for cleaving the G protein in Vero cells and demonstrated that cleavage likely occurs during endocytic recycling. Others have shown that the Nipah virus fusion protein is activated by cathepsin L during recycling (25,26). In addition, endocytic recycling is important in the envelopment of RSV (27,28).…”

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confidence: 99%

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Corry

Johnson

Cornwell

et al. 2016

J Virol

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All live attenuated respiratory syncytial virus (RSV) vaccines that have advanced to clinical trials have been produced in Vero cells. The attachment (G) glycoprotein in virions produced in these cells is smaller than that produced in other immortalized cells due to cleavage. These virions are 5-fold less infectious for primary well-differentiated human airway epithelial (HAE) cell cultures. Because HAE cells are isolated directly from human airways, Vero cell-grown vaccine virus would very likely be similarly inefficient at initiating infection of the nasal epithelium following vaccination, and therefore, a larger inoculum would be required for effective vaccination. We hypothesized that Vero cell-derived virus containing an intact G protein would be more infectious for HAE cell cultures. Using protease inhibitors with increasing specificity, we identified cathepsin L to be the protease responsible for cleavage. Our evidence suggests that cleavage occurs in the late endosome or lysosome during endocytic recycling. Cathepsin L activity was 100-fold greater in Vero cells than in HeLa cells. In addition, cathepsin L was able to cleave the G protein in Vero cell-grown virions but not in HeLa cell-grown virions, suggesting a difference in G-protein posttranslational modification in the two cell lines. We identified by mutagenesis amino acids important for cleavage, and these amino acids included a likely cathepsin L cleavage site. Virus containing a modified, noncleavable G protein produced in Vero cells was 5-fold more infectious for HAE cells in culture, confirming our hypothesis and indicating the value of including such a mutation in future live attenuated RSV vaccines. IMPORTANCEWorldwide, RSV is the second leading infectious cause of infant death, but no vaccine is available. Experimental live attenuated RSV vaccines are grown in Vero cells, but during production the virion attachment (G) glycoprotein is cleaved. Virions containing a cleaved G protein are less infectious for primary airway epithelial cells, the natural RSV target. In the study described here we identified the protease responsible, located the cleavage site, and demonstrated that cleavage likely occurs during endocytic recycling. Moreover, we showed that the infectivity of Vero cell-derived virus for primary airway epithelial cells is increased 5-fold if the virus contains a mutation in the G protein that prevents cleavage. The blocking of cleavage should improve RSV vaccine yield, consequently reducing production costs. Posttranslational cleavage of the fusion glycoprotein of many viruses plays an essential role in activation; however, cleavage of the RSV G protein is a novel example of a detrimental effect of cleavage on virus infectivity. In adults and teenagers, most respiratory syncytial virus (RSV) infections produce upper respiratory tract infection and symptoms. However, in infants, the immunocompromised, and the elderly, RSV has the potential to cause life-threatening lower respiratory tract infection (1-4). Worldwide, in 2010 ...

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Nipah virus fusion protein: Influence of cleavage site mutations on the cleavability by cathepsin L, trypsin and furin

Cited by 11 publications

References 49 publications

Activation of the Nipah Virus Fusion Protein in MDCK Cells Is Mediated by Cathepsin B within the Endosome-Recycling Compartment

Activation of the Nipah Virus Fusion Protein in MDCK Cells Is Mediated by Cathepsin B within the Endosome-Recycling Compartment

Cell Entry of Enveloped Viruses

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

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