The role of single N-glycans in proteolytic processing and cell surface transport of the Lassa virus glycoprotein GP-C

Eichler, Robert; Lenz, Oliver; Garten, Wolfgang; Strecker, Thomas

doi:10.1186/1743-422x-3-41

Cited by 67 publications

(61 citation statements)

References 27 publications

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“…Arenavirus GP N-linked glycans play significant biochemical roles in virus-cell interactions (34,35). Previous studies found that specific N-linked glycans in LCMV GP1 were necessary for GP trafficking, fusion activity, and infectivity (34).…”

Section: Resultsmentioning

confidence: 99%

“…LASV GP1 contains seven conserved N-linked glycosylation sites. Removing glycans at positions 81, 91, 101, and 121 was previously shown to inhibit GPC processing into GP1 and GP2 (35), although receptor binding and fusion activity were not evaluated. We reproduced the seven N-glycan mutants by changing glycosylation site motifs from N-X-S/T to N-X-A.…”

Section: Resultsmentioning

confidence: 99%

“…However, we found that the remaining five N-glycan mutations resulted in detectable levels of GP1-GP2 processing, including T101 and S121. Our use of a codon-optimized expression construct and examining surface material 36 h after transfection, rather than 24 h, may have enabled T101A and S121A mutants to reach detectable steady-state levels of processed GP1-GP2 at the cell surface (35).…”

Section: Resultsmentioning

confidence: 99%

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Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions Required for Alpha-Dystroglycan Utilization

Acciani

Alston

Zhao

et al. 2017

J Virol

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Lassa virus (LASV) is an enveloped RNA virus endemic to West Africa and responsible for severe cases of hemorrhagic fever. Virus entry is mediated by the glycoprotein complex consisting of a stable-signal peptide, a receptor-binding subunit, GP1, and a viral-host membrane fusion subunit, GP2. Several cellular receptors can interact with the GP1 subunit and mediate viral entry, including alpha-dystroglycan (␣DG) and lysosome-associated membrane protein 1 (LAMP1). In order to define the regions within GP1 that interact with the cellular receptors, we implemented insertional mutagenesis, carbohydrate shielding, and alanine scanning mutagenesis. Eighty GP constructs were engineered and evaluated for GP1-GP2 processing, surface expression, and the ability to mediate cell-to-cell fusion after low-pH exposure. To examine virus-to-cell entry, 49 constructs were incorporated onto vesicular stomatitis virus (VSV) pseudoparticles and transduction efficiencies were monitored in HAP1 and HAP1-ΔDAG1 cells that differentially produce the ␣DG cell surface receptor. Seven constructs retained efficient transduction in HAP1-ΔDAG1 cells yet poorly transduced HAP1 cells, suggesting that they are involved in ␣DG utilization. Residues H141, N146, F147, and Y150 cluster at the predicted central core of the trimeric interface and are important for GP-␣DG interaction. Additionally, H92A-H93A, 150HA, 172HA, and 230HA displayed reduced transduction in both HAP1 and HAP1-ΔDAG1 cells, despite efficient cell-to-cell fusion activity. These mutations may interfere with interactions with the endosomal receptor LAMP1 or interfere at another stage in entry that is common to both cell lines. Insight gained from these data can aid in the development of more-effective entry inhibitors by blocking receptor interactions.IMPORTANCE Countries in which Lassa virus is endemic, such as Nigeria, Sierra Leone, Guinea, and Liberia, usually experience a seasonal outbreak of the virus from December to March. Currently, there is neither a preventative vaccine nor a therapeutic available to effectively treat severe Lassa fever. One way to thwart virus infection is to inhibit interaction with cellular receptors. It is known that the GP1 subunit of the Lassa glycoprotein complex plays a critical role in receptor recognition. Our results highlight a region within the Lassa virus GP1 protein that interacts with the cellular receptor alpha-dystroglycan. This information may be used for future development of new Lassa virus antivirals.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions Required for Alpha-Dystroglycan Utilization

Acciani

Alston

Zhao

et al. 2017

J Virol

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“…The LASV GPC contains eleven potential N-glycosylation sites, but disruption of only six of them affected GPC processing indicating that these N-glycans were essential for correct conformation of GPC in order to be cleaved by the host cell subtilase [168]. The LCMV GPC contains eight potentially glycosylated sites [169].…”

Section: The Lassa Virus Signal Peptide and Gpc Processingmentioning

confidence: 99%

Lassa Virus Genome

Igor¹,

Sica²

2006

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Lassa virus (LASV), the most dangerous human pathogen among the Arenaviridae, belongs to a complex of genetically related virus strains responsible for the deaths of thousands of people in West Africa each year. The virus has a bi-segmented (L and S) single-stranded RNA genome. Each segment contains two genes in ambisense orientation. The L RNA encodes a large protein, L, or RdRp and a small zinc-binding, Z protein. The S RNA encodes the major structural proteins, nucleoprotein (NP) and glycoprotein precursor (GPC), cleaved into signal peptide, GP1, and GP2 glycoproteins. Genetic diversity among LASV strains is the highest within the family Arenaviridae and NP and RdRp genes are the most variable among LASV genes. The LASV genetic diversity is a great challenge for vaccine development.In addition to LASV and the prototype lymphocytic choriomeningitis virus (LCMV), the Old World group of arenaviruses includes three other related viruses, Mopeia (MOPV), Mobala (MOBV), and Ippy (IPPYV). These viruses as well as a MOP/LAS reassortant carrying the L RNA segment from MOPV and S RNA segment from LASV are non-pathogenic for experimental animals and are able to induce protective immunity against LASV. Lassa Fever pathogenesis is a sum of the effects induced by viral replication and immune response. The goal of this review is to cover recent publications on viral and host genes that control LASV virulence. The full-length genome sequence of LASV isolates and LASV-related nonpathogenic arenaviruses will provide a useful genetic tool to map LASV genes involved in virulence and to gain insight into phylogeny and evolution of the Old World arenaviruses.

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“…гликопротеины G1 и G2 содержат несколько участков гликозилирования, которые являются необ-ходимыми для осуществления белками своих функ-ций [18]. в частности, гликопротеин вируса ласса содержит шесть участков гликозилирования [5].…”

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Molecular-Biological Peculiarities of Arenavirus Reproduction in Sensitive Cells

Сизикова¹,

Лебедев²,

Пантюхов³

et al. 2017

Problemy Osobo Opasnykh Infektsii

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The role of single N-glycans in proteolytic processing and cell surface transport of the Lassa virus glycoprotein GP-C

Cited by 67 publications

References 27 publications

Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions Required for Alpha-Dystroglycan Utilization

Mutational Analysis of Lassa Virus Glycoprotein Highlights Regions Required for Alpha-Dystroglycan Utilization

Lassa Virus Genome

Molecular-Biological Peculiarities of Arenavirus Reproduction in Sensitive Cells

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