In Vitro reconstitution reveals key intermediate states of trimer formation by the dengue virus membrane fusion protein

Maofu Liao, Claudia Sánchez-San Martín, Aihua Zheng, Margaret Kielian

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The flavivirus dengue virus (DV) infects cells through a low-pH-triggered membrane fusion reaction mediated by the viral envelope protein E. E is an elongated transmembrane protein with three domains and is organized as a homodimer on the mature virus particle. During fusion, the E protein homodimer dissociates, inserts the hydrophobic fusion loop into target membranes, and refolds into a trimeric hairpin in which domain III (DIII) packs against the central trimer. It is clear that E refolding drives membrane fusion, but the steps in hairpin formation and their pH requirements are unclear. Here, we have used truncated forms of the DV E protein to reconstitute trimerization in vitro. Protein constructs containing domains I and II (DI/II) were monomeric and interacted with membranes to form core trimers. DI/II-membrane interaction and trimerization occurred efficiently at both neutral and low pH. The DI/II core trimer was relatively unstable and could be stabilized by binding exogenous DIII or by the formation of mixed trimers containing DI/II plus E protein with all three domains. The mixed trimer had unoccupied DIII interaction sites that could specifically bind exogenous DIII at either low or neutral pH. Truncated DV E proteins thus reconstitute hairpin formation and define properties of key domain interactions during DV fusion.

Original languageEnglish (US)
Pages (from-to)5730-5740
Number of pages11
JournalJournal of Virology
Volume84
Issue number11
DOIs
StatePublished - Jun 2010

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Membrane Fusion Proteins
Viral Fusion Proteins
Virus Internalization
Dengue virus
Dengue Virus
Proteins
Membrane Fusion
proteins
Membranes
Flavivirus
transmembrane proteins
Viral Envelope Proteins
virion
Virion
In Vitro Techniques
membrane fusion

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

In Vitro reconstitution reveals key intermediate states of trimer formation by the dengue virus membrane fusion protein. / Liao, Maofu; Sánchez-San Martín, Claudia; Zheng, Aihua; Kielian, Margaret.

In: Journal of Virology, Vol. 84, No. 11, 06.2010, p. 5730-5740.

Research output: Contribution to journalArticle

Liao, Maofu ; Sánchez-San Martín, Claudia ; Zheng, Aihua ; Kielian, Margaret. / In Vitro reconstitution reveals key intermediate states of trimer formation by the dengue virus membrane fusion protein. In: Journal of Virology. 2010 ; Vol. 84, No. 11. pp. 5730-5740.
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