Niemann-Pick C1 (NPC1)/NPC1-like1 chimeras define sequences critical for NPC1's function as a filovirus entry receptor

Anuja Krishnan, Emily Happy Miller, Andrew S. Herbert, Melinda Ng, Esther Ndungo, Sean P. Whelan, John M. Dye, Kartik Chandran

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

We recently demonstrated that Niemann-Pick C1 (NPC1), a ubiquitous 13-pass cellular membrane protein involved in lysosomal cholesterol transport, is a critical entry receptor for filoviruses. Here we show that Niemann-Pick C1-like1 (NPC1L1), an NPC1 paralog and hepatitis C virus entry factor, lacks filovirus receptor activity. We exploited the structural similarity between NPC1 and NPC1L1 to construct and analyze a panel of chimeras in which NPC1L1 sequences were replaced with cognate sequences from NPC1. Only one chimera, NPC1L1 containing the second luminal domain (C) of NPC1 in place of its own, bound to the viral glycoprotein, GP. This engineered protein mediated authentic filovirus infection nearly as well as wild-type NPC1, and more efficiently than did a minimal NPC1 domain C-based receptor recently described by us. A reciprocal chimera, NPC1 containing NPC1L1's domain C, was completely inactive. Remarkably, an intra-domain NPC1L1-NPC1 chimera bearing only a ~130-amino acid N-terminal region of NPC1 domain C could confer substantial viral receptor activity on NPC1L1. Taken together, these findings account for the failure of NPC1L1 to serve as a filovirus receptor, highlight the central role of the luminal domain C of NPC1 in filovirus entry, and reveal the direct involvement of N-terminal domain C sequences in NPC1's function as a filovirus receptor.

Original languageEnglish (US)
Pages (from-to)2471-2484
Number of pages14
JournalViruses
Volume4
Issue number11
DOIs
StatePublished - Nov 2012

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Virus Internalization
Hepacivirus
Glycoproteins
Membrane Proteins
Cholesterol
Amino Acids
Infection
Proteins

Keywords

  • Ebola virus
  • Filovirus
  • Host factor
  • Marburg virus
  • Niemann-Pick C1
  • Niemann-Pick C1-like1
  • NPC1
  • NPC1L1
  • Viral entry
  • Viral receptor

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

Niemann-Pick C1 (NPC1)/NPC1-like1 chimeras define sequences critical for NPC1's function as a filovirus entry receptor. / Krishnan, Anuja; Miller, Emily Happy; Herbert, Andrew S.; Ng, Melinda; Ndungo, Esther; Whelan, Sean P.; Dye, John M.; Chandran, Kartik.

In: Viruses, Vol. 4, No. 11, 11.2012, p. 2471-2484.

Research output: Contribution to journalArticle

Krishnan, A, Miller, EH, Herbert, AS, Ng, M, Ndungo, E, Whelan, SP, Dye, JM & Chandran, K 2012, 'Niemann-Pick C1 (NPC1)/NPC1-like1 chimeras define sequences critical for NPC1's function as a filovirus entry receptor', Viruses, vol. 4, no. 11, pp. 2471-2484. https://doi.org/10.3390/v4112471
Krishnan, Anuja ; Miller, Emily Happy ; Herbert, Andrew S. ; Ng, Melinda ; Ndungo, Esther ; Whelan, Sean P. ; Dye, John M. ; Chandran, Kartik. / Niemann-Pick C1 (NPC1)/NPC1-like1 chimeras define sequences critical for NPC1's function as a filovirus entry receptor. In: Viruses. 2012 ; Vol. 4, No. 11. pp. 2471-2484.
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