Over-expression of mammalian sialidase NEU3 reduces Newcastle disease virus entry and propagation in COS7 cells

• Published in: Biochimica et biophysica acta Vol. 1780; no. 3; pp. 504 - 512
• Main Authors: Anastasia, Luigi, Holguera, Javier, Bianchi, Anna, D'Avila, Francesca, Papini, Nadia, Tringali, Cristina, Monti, Eugenio, Villar, Enrique, Venerando, Bruno, Muñoz-Barroso, Isabel, Tettamanti, Guido
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2008
• Subjects: Animals; Blotting, Western; Cell Fusion; Cell Membrane - virology; Cercopithecus aethiops; Chromatography, Thin Layer; COS Cells; COS7 cell; G(M1) Ganglioside - analogs & derivatives; G(M1) Ganglioside - metabolism; GD1a ganglioside; Gene Expression; Giant Cells - virology; HN Protein - metabolism; Lipid raft; NDV; Neuraminidase - genetics; Neuraminidase - metabolism; Newcastle Disease - virology; Newcastle disease virus - physiology; Sialidase NEU3; Virus Internalization; Virus Replication; PBS; NDV; GD1a ganglioside; Lipid raft; COS7 cell; F; HN; MOI; Neu5Ac; F0; GFP; DMEM; DRMs; Sialidase NEU3; FBS; R18; MmNEU3; pfu; HPTLC
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2007.11.011

The paramyxovirus Newcastle Disease Virus (NDV) binds to sialic acid-containing glycoconjugates, sialoglycoproteins and sialoglycolipids (gangliosides) of host cell plasma membrane through its hemagglutinin-neuraminidase (sialidase) HN glycoprotein. We hypothesized that the modifications of the cell surface ganglioside pattern determined by over-expression of the mammalian plasma-membrane associated, ganglioside specific, sialidase NEU3 would affect the virus-host cell interactions. Using COS7 cells as a model system, we observed that over-expression of the murine MmNEU3 did not affect NDV binding but caused a marked reduction in NDV infection and virus propagation through cell–cell fusion. Moreover, since GD1a was greatly reduced in COS7 cells following NEU3-over-expression, we added [ 3H]-labelled GD1a to COS7 cells under conditions that block intralysosomal metabolic processing, and we observed a marked increase of GD1a cleavage to GM1 during NDV infection, indicating a direct involvement of the virus sialidase and host cell GD1a in NDV infectivity. Therefore, the decrease of GD1a in COS7 cell membrane upon MmNEU3 over-expression is likely to be instrumental to NDV reduced infection. Evidence was also provided for the preferential association of NDV-HN at 4 °C to detergent resistant microdomains (DRMs) of COS7 cells plasma membranes.