Abstract
Cells genetically deficient in sphingomyelin synthase-1 (SGMS1) or blocked in their synthesis pharmacologically through exposure to a serine palmitoyltransferase inhibitor (myriocin) show strongly reduced surface display of influenza virus glycoproteins hemag-glutinin (HA) and neuraminidase (NA). The transport of HA to the cell surface was assessed by accessibility of HA on intact cells to exogenously added trypsin and to HA-specific antibodies. Rates of de novo synthesis of viral proteins in wild-type and SGMS1-deficient cells were equivalent, and HA negotiated the intracellular trafficking pathway through the Golgi normally. We engineered a strain of influenza virus to allow site-specific labeling of HA and NA using sortase. Accessibility of both HA and NA to sortase was blocked in SGMS1-deficient cells and in cells exposed to myriocin, with a corresponding inhibition of the release of virus particles from infected cells. Generation of influenza virus particles thus critically relies on a functional sphingomyelin biosynthetic pathway, required to drive influenza viral glycoproteins into lipid domains of a composition compatible with virus budding and release.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6406-6411 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 110 |
| Issue number | 16 |
| DOIs | |
| State | Published - Apr 16 2013 |
| Externally published | Yes |
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Intact sphingomyelin biosynthetic pathway is essential for intracellular transport of influenza virus glycoproteins. / Tafesse, Fikadu; Sanyal, Sumana; Ashour, Joseph; Guimaraes, Carla P.; Hermansson, Martin; Somerharju, Pentti; Ploegh, Hidde L.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, No. 16, 16.04.2013, p. 6406-6411.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Intact sphingomyelin biosynthetic pathway is essential for intracellular transport of influenza virus glycoproteins
AU - Tafesse, Fikadu
AU - Sanyal, Sumana
AU - Ashour, Joseph
AU - Guimaraes, Carla P.
AU - Hermansson, Martin
AU - Somerharju, Pentti
AU - Ploegh, Hidde L.
PY - 2013/4/16
Y1 - 2013/4/16
N2 - Cells genetically deficient in sphingomyelin synthase-1 (SGMS1) or blocked in their synthesis pharmacologically through exposure to a serine palmitoyltransferase inhibitor (myriocin) show strongly reduced surface display of influenza virus glycoproteins hemag-glutinin (HA) and neuraminidase (NA). The transport of HA to the cell surface was assessed by accessibility of HA on intact cells to exogenously added trypsin and to HA-specific antibodies. Rates of de novo synthesis of viral proteins in wild-type and SGMS1-deficient cells were equivalent, and HA negotiated the intracellular trafficking pathway through the Golgi normally. We engineered a strain of influenza virus to allow site-specific labeling of HA and NA using sortase. Accessibility of both HA and NA to sortase was blocked in SGMS1-deficient cells and in cells exposed to myriocin, with a corresponding inhibition of the release of virus particles from infected cells. Generation of influenza virus particles thus critically relies on a functional sphingomyelin biosynthetic pathway, required to drive influenza viral glycoproteins into lipid domains of a composition compatible with virus budding and release.
AB - Cells genetically deficient in sphingomyelin synthase-1 (SGMS1) or blocked in their synthesis pharmacologically through exposure to a serine palmitoyltransferase inhibitor (myriocin) show strongly reduced surface display of influenza virus glycoproteins hemag-glutinin (HA) and neuraminidase (NA). The transport of HA to the cell surface was assessed by accessibility of HA on intact cells to exogenously added trypsin and to HA-specific antibodies. Rates of de novo synthesis of viral proteins in wild-type and SGMS1-deficient cells were equivalent, and HA negotiated the intracellular trafficking pathway through the Golgi normally. We engineered a strain of influenza virus to allow site-specific labeling of HA and NA using sortase. Accessibility of both HA and NA to sortase was blocked in SGMS1-deficient cells and in cells exposed to myriocin, with a corresponding inhibition of the release of virus particles from infected cells. Generation of influenza virus particles thus critically relies on a functional sphingomyelin biosynthetic pathway, required to drive influenza viral glycoproteins into lipid domains of a composition compatible with virus budding and release.
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UR - http://www.scopus.com/inward/citedby.url?scp=84876276496&partnerID=8YFLogxK
U2 - 10.1073/pnas.1219909110
DO - 10.1073/pnas.1219909110
M3 - Article
C2 - 23576732
AN - SCOPUS:84876276496
VL - 110
SP - 6406
EP - 6411
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 16
ER -