Enhancing glycoprotein sialylation by targeted gene silencing in mammalian cells

Zhang, Min; Koskie, Kerry; Ross, J. S.; Kayser, Kevin J.; Caple, Matthew V.

doi:10.1002/bit.22633

Cited by 46 publications

(31 citation statements)

References 43 publications

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“…NEU3 was designated the plasma membrane-associated sialidase (36,40). When a cDNA for human NEU3 was transfected into COS-7 cells, the protein was immunolocalized to the plasma membrane (40,71). The murine and bovine enzymes similarly associate with the plasma membrane (36,45,70).…”

Section: Discussionmentioning

confidence: 99%

Human airway epithelia express catalytically active NEU3 sialidase

Lillehoj

Hyun²,

Feng

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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acids on glycoconjugates play a pivotal role in many biological processes. In the airways, sialylated glycoproteins and glycolipids are strategically positioned on the plasma membranes of epithelia to regulate receptor-ligand, cell-cell, and host-pathogen interactions at the molecular level. We now demonstrate, for the first time, sialidase activity for ganglioside substrates in human airway epithelia. Of the four known mammalian sialidases, NEU3 has a substrate preference for gangliosides and is expressed at mRNA and protein levels at comparable abundance in epithelia derived from human trachea, bronchi, small airways, and alveoli. In small airway and alveolar epithelia, NEU3 protein was immunolocalized to the plasma membrane, cytosolic, and nuclear subcellular fractions. Small interfering RNA-induced silencing of NEU3 expression diminished sialidase activity for a ganglioside substrate by Ͼ70%. NEU3 immunostaining of intact human lung tissue could be localized to the superficial epithelia, including the ciliated brush border, as well as to nuclei. However, NEU3 was reduced in subepithelial tissues. These results indicate that human airway epithelia express catalytically active NEU3 sialidase.

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Section: Discussionmentioning

confidence: 99%

Human airway epithelia express catalytically active NEU3 sialidase

Lillehoj

Hyun²,

Feng

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…The sialidases are poorly investigated, but some experimental data exists for human membrane neuraminidase hNeu3 ( Monti et al , 2000; Zhang et al , 2010) and the enzyme was selected for cloning.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of defense strategy against Influenza A in cell line models

et al. 2018

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Influenza virus can cause both seasonal infections and unpredictable pandemics. Rapidly evolving avian H5N1 virus is getting increasingly infective for humans. Since avian Influenza can be transmitted by domestic birds, serving as a key link between wild aquatic birds and humans, an effective measure to control the influenza transmission would be eradication of the infection in poultry. It is known that the virus penetrates into the cell through binding with the terminal oligosaccharides - sialic acids (SA) - on the cell surfaces. Removal of SA might be a potential antiviral strategy. An approach to developing chicken lines that are resistant to influenza viruses could be the creation of genetically modified birds. Thus it is necessary to select a gene that provides defense to influenza. Here we have expressed in cells a range of exogenous sialidases and estimated their activity and specificity towards SA residues. Several bacterial, viral and human sialidases were tested. We adopted bacterial sialidases from and for expression on the cell surface by fusing catalytic domains with transmembrane domains. We also selected Influenza A/PuertoRico/8/34/H1N1 neuraminidase and human membrane sialidase () genes. Lectin binding assay was used for estimation of a α (2,3)-sialylation level by fluorescent microscopy and FACS. We compared sialidases from bacteria, Influenza virus and human. Sialidases from and Influenza A neuraminidase effectively cleaved α (2-3)-SA receptors. Viral neuraminidase demonstrated a higher activity. Sialidases from and did not show any activity against α (2-3) SA under physiological conditions. : Our results demonstrated that sialidases with different specificity and activity can be selected as genes providing antiviral defence. Combining chosen sialidases with different activity together with tissue-specific promoters would provide an optimal level of desialilation to prevent infection. Tissue specific expression of the sialidases could protect domestic birds from infection.

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“…However, this knockdown of Neu2 did not increase the total sialylation. To resolve this problem, Zhang et al conducted a similar experiment and knocked down both Neu2 and Neu3 gene using shRNA [30]. The result indicated the simultaneous knockdown of both, Neu2 and Neu3, could moderately increase the sialylation.…”

Section: Knockdown Of Sialidasesmentioning

confidence: 90%