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Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis
10:23

Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis

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Effect of host cell lipid metabolism on alphavirus replication, virion morphogenesis, and infectivity

Ching G Ng1, Isabelle Coppens, Dhanasekaran Govindarajan

  • 1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.

Proceedings of the National Academy of Sciences of the United States of America
|October 11, 2008
PubMed

Insights

Acid sphingomyelinase deficiency enhances Sindbis virus (SINV) replication and infectivity. This lipid metabolism disorder leads to more infectious virions, suggesting a new viral control mechanism.

Area of Science:

  • Virology
  • Cell Biology
  • Lipid Metabolism

Background:

  • Sindbis virus (SINV) causes encephalomyelitis.
  • Lipid membranes, especially cholesterol and sphingomyelin (SM), are crucial for viral lifecycle.
  • Acid sphingomyelinase (ASMase) regulates SM levels; its deficiency causes Niemann-Pick disease type A (NPD-A).

Purpose of the Study:

  • To elucidate mechanisms by which ASMase deficiency enhances SINV replication.
  • To compare SINV replication and pathogenesis in normal human fibroblasts (NHF) and NPD-A fibroblasts (NPAF).

Main Methods:

  • Comparison of SINV replication in NPAF and NHF.
  • Analysis of viral RNA, protein synthesis, and virion characteristics.
  • Assessment of cellular lipid accumulation in NPAF.

Main Results:

  • NPAF accumulated cholesterol- and sphingolipid-rich late endosomes/lysosomes.
  • SINV replicated faster and to higher titers in NPAF.
  • Virions budding from NPAF were significantly more infectious and morphologically distinct from those from NHF.

Conclusions:

  • ASMase deficiency enhances SINV replication and pathogenesis.
  • Cellular lipid composition influences alphavirus morphogenesis and virion infectivity.
  • Cellular regulation of alphavirus morphogenesis is a novel mechanism for controlling virus replication and spread.

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