Premium
Direct Imaging of the Cholesterol and Sphingolipid Abundance at the Site of Influenza Virus Assembly with High‐Resolution SIMS
Author(s) -
Kraft Mary L,
Yeager Ashley N,
Weber Peter K,
Joshua Joshua
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.797.5
Subject(s) - sphingolipid , virus , hemagglutinin (influenza) , cholesterol , budding , membrane , lipid raft , biology , influenza a virus , virology , microbiology and biotechnology , chemistry , biochemistry
The influenza virus is hypothesized to assemble and bud from plasma membrane domains that are enriched with cholesterol and sphingolipids. This hypothesis is consistent with the reported enrichment of the influenza viral envelope with cholesterol and sphingolipids, the reduction in influenza virus protein clustering in the plasma membrane induced by cholesterol depletion, and the sensitivity of influenza virus formation to cellular cholesterol levels. Nonetheless, the levels of cholesterol and sphingolipids in the plasma membrane at the site of influenza virus budding have never been directly measured. The main challenge has been a lack of analytical methods with the sensitivity and lateral resolution that is required to measure the relative cholesterol and sphingolipid levels in the plasma membrane on the 100‐nm‐length scales that are relevant to influenza virus budding. We recently overcame this challenge by using a secondary ion mass spectrometry (SIMS) technique with high spatial resolution and sensitivity to image the distributions of stable isotope‐labeled cholesterol, sphingolipids, and immunolabeled influenza hemagglutinin in the plasma membranes of virus‐infected MDCK cells. Our results show that the plasma membranes of both influenza‐infected and uninfected MDCK cells contained regions that were enriched with sphingolipids, but cholesterol appeared to be uniformly distributed within the membrane. Examination of the influenza‐infected MDCK cells revealed the hemagglutinin‐rich sites of influenza virus assembly in the plasma membrane were not enriched with cholesterol, and seldom enriched with sphingolipids. These direct imaging results demonstrate that the influenza virus does not assemble within plasma membrane domains that are enriched with cholesterol and sphingolipids. Support or Funding Information This research was supported by the U.S. National Science Foundation under CHE 15‐08662. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .