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Sialic acid-dependent cell entry of human enterovirus D68.

Liu Y, Sheng J, Baggen J, Meng G, Xiao C, Thibaut HJ, van Kuppeveld FJ, Rossmann MG - Nat Commun (2015)

Bottom Line: Human enterovirus D68 (EV-D68) is a causative agent of childhood respiratory diseases and has now emerged as a global public health threat.Nevertheless, knowledge of the tissue tropism and pathogenesis of EV-D68 has been hindered by a lack of studies on the receptor-mediated EV-D68 entry into host cells.Crystal structures of EV-D68 in complex with sialylated glycan receptor analogues show that they bind into the 'canyon' on the virus surface.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Hockmeyer Hall of Structural Biology, 240 South Martin Jischke Drive, Purdue University, West Lafayette, Indiana 47907, USA.

ABSTRACT
Human enterovirus D68 (EV-D68) is a causative agent of childhood respiratory diseases and has now emerged as a global public health threat. Nevertheless, knowledge of the tissue tropism and pathogenesis of EV-D68 has been hindered by a lack of studies on the receptor-mediated EV-D68 entry into host cells. Here we demonstrate that cell surface sialic acid is essential for EV-D68 to bind to and infect susceptible cells. Crystal structures of EV-D68 in complex with sialylated glycan receptor analogues show that they bind into the 'canyon' on the virus surface. The sialic acid receptor induces a cascade of conformational changes in the virus to eject a fatty-acid-like molecule that regulates the stability of the virus. Thus, virus binding to a sialic acid receptor and to immunoglobulin-like receptors used by most other enteroviruses share a conserved mechanism for priming viral uncoating and facilitating cell entry.

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Cell surface sialic acid has a crucial role in EV-D68 (strain Fermon CA62-1) attachment and infection.(a–c) Growth curve of EV-D68 in susceptible cell lines cells. Neuraminidase treatment of susceptible cell lines inhibits EV-D68 infection. (d) HAP1 cells (WT and CMAS-KO) were infected with EV-D68 at an MOI of 1 and virus titres were determined at 0 h (T=0) or 10-h post infection. (e,f) Histograms showing virus binding to susceptible cells. P<0.0001 by Student's t-test. (g) Preincubation of 6′SL, 6′SLN or 3′SLN with EV-D68 prevents killing of RD cells caused by virus infection. (h) Preincubation of 6′SLN or 3′SLN with EV-D68 inhibits viral attachment to RD cells. All data are represented as mean±s.d. Experiments were performed at least in triplicate.
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f1: Cell surface sialic acid has a crucial role in EV-D68 (strain Fermon CA62-1) attachment and infection.(a–c) Growth curve of EV-D68 in susceptible cell lines cells. Neuraminidase treatment of susceptible cell lines inhibits EV-D68 infection. (d) HAP1 cells (WT and CMAS-KO) were infected with EV-D68 at an MOI of 1 and virus titres were determined at 0 h (T=0) or 10-h post infection. (e,f) Histograms showing virus binding to susceptible cells. P<0.0001 by Student's t-test. (g) Preincubation of 6′SL, 6′SLN or 3′SLN with EV-D68 prevents killing of RD cells caused by virus infection. (h) Preincubation of 6′SLN or 3′SLN with EV-D68 inhibits viral attachment to RD cells. All data are represented as mean±s.d. Experiments were performed at least in triplicate.

Mentions: The possible use of sialic acid as a cellular receptor by EV-D68 was examined by performing attachment and infectivity assays. We found that removal of cell surface sialic acid by neuraminidase treatment of HeLa cells, human rhabdomyosarcoma (RD) cells and human lung embryonic fibroblast (HELF) cells significantly reduced infectivity of the EV-D68 Fermon prototype strain (Fig. 1a–c). These results suggest that sialic acid might be a functional receptor in these cell lines. To obtain further evidence that sialic acid is a functional receptor for EV-D68, human HAP1 cells were used in which the sialic acid activating enzyme cytidine monophosphate N-acetylneuraminic acid synthase (CMAS) had been knocked out. These cells are devoid of sialic acids on their surface and had previously been shown to be highly resistant to influenza A virus infection30. We found that these cells were resistant to EV-D68 (Fermon strain) infection, further confirming the importance of sialic acid for EV-D68 infection (Fig. 1d). Furthermore, neuraminidase treatment of RD and HELF cells led to significantly decreased virus attachment (Fig. 1e,f).


Sialic acid-dependent cell entry of human enterovirus D68.

Liu Y, Sheng J, Baggen J, Meng G, Xiao C, Thibaut HJ, van Kuppeveld FJ, Rossmann MG - Nat Commun (2015)

Cell surface sialic acid has a crucial role in EV-D68 (strain Fermon CA62-1) attachment and infection.(a–c) Growth curve of EV-D68 in susceptible cell lines cells. Neuraminidase treatment of susceptible cell lines inhibits EV-D68 infection. (d) HAP1 cells (WT and CMAS-KO) were infected with EV-D68 at an MOI of 1 and virus titres were determined at 0 h (T=0) or 10-h post infection. (e,f) Histograms showing virus binding to susceptible cells. P<0.0001 by Student's t-test. (g) Preincubation of 6′SL, 6′SLN or 3′SLN with EV-D68 prevents killing of RD cells caused by virus infection. (h) Preincubation of 6′SLN or 3′SLN with EV-D68 inhibits viral attachment to RD cells. All data are represented as mean±s.d. Experiments were performed at least in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4660200&req=5

f1: Cell surface sialic acid has a crucial role in EV-D68 (strain Fermon CA62-1) attachment and infection.(a–c) Growth curve of EV-D68 in susceptible cell lines cells. Neuraminidase treatment of susceptible cell lines inhibits EV-D68 infection. (d) HAP1 cells (WT and CMAS-KO) were infected with EV-D68 at an MOI of 1 and virus titres were determined at 0 h (T=0) or 10-h post infection. (e,f) Histograms showing virus binding to susceptible cells. P<0.0001 by Student's t-test. (g) Preincubation of 6′SL, 6′SLN or 3′SLN with EV-D68 prevents killing of RD cells caused by virus infection. (h) Preincubation of 6′SLN or 3′SLN with EV-D68 inhibits viral attachment to RD cells. All data are represented as mean±s.d. Experiments were performed at least in triplicate.
Mentions: The possible use of sialic acid as a cellular receptor by EV-D68 was examined by performing attachment and infectivity assays. We found that removal of cell surface sialic acid by neuraminidase treatment of HeLa cells, human rhabdomyosarcoma (RD) cells and human lung embryonic fibroblast (HELF) cells significantly reduced infectivity of the EV-D68 Fermon prototype strain (Fig. 1a–c). These results suggest that sialic acid might be a functional receptor in these cell lines. To obtain further evidence that sialic acid is a functional receptor for EV-D68, human HAP1 cells were used in which the sialic acid activating enzyme cytidine monophosphate N-acetylneuraminic acid synthase (CMAS) had been knocked out. These cells are devoid of sialic acids on their surface and had previously been shown to be highly resistant to influenza A virus infection30. We found that these cells were resistant to EV-D68 (Fermon strain) infection, further confirming the importance of sialic acid for EV-D68 infection (Fig. 1d). Furthermore, neuraminidase treatment of RD and HELF cells led to significantly decreased virus attachment (Fig. 1e,f).

Bottom Line: Human enterovirus D68 (EV-D68) is a causative agent of childhood respiratory diseases and has now emerged as a global public health threat.Nevertheless, knowledge of the tissue tropism and pathogenesis of EV-D68 has been hindered by a lack of studies on the receptor-mediated EV-D68 entry into host cells.Crystal structures of EV-D68 in complex with sialylated glycan receptor analogues show that they bind into the 'canyon' on the virus surface.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Hockmeyer Hall of Structural Biology, 240 South Martin Jischke Drive, Purdue University, West Lafayette, Indiana 47907, USA.

ABSTRACT
Human enterovirus D68 (EV-D68) is a causative agent of childhood respiratory diseases and has now emerged as a global public health threat. Nevertheless, knowledge of the tissue tropism and pathogenesis of EV-D68 has been hindered by a lack of studies on the receptor-mediated EV-D68 entry into host cells. Here we demonstrate that cell surface sialic acid is essential for EV-D68 to bind to and infect susceptible cells. Crystal structures of EV-D68 in complex with sialylated glycan receptor analogues show that they bind into the 'canyon' on the virus surface. The sialic acid receptor induces a cascade of conformational changes in the virus to eject a fatty-acid-like molecule that regulates the stability of the virus. Thus, virus binding to a sialic acid receptor and to immunoglobulin-like receptors used by most other enteroviruses share a conserved mechanism for priming viral uncoating and facilitating cell entry.

Show MeSH
Related in: MedlinePlus