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Structure of the three-way helical junction of the hepatitis C virus IRES element

Structure of the three-way helical junction of the hepatitis C virus IRES element

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Authors

  • Jonathan Ouellet
  • Sonya Melcher
  • Asif Iqbal
  • Yiliang Ding
  • David M. J. Lilley

Research units

Info

Original languageEnglish
Pages1597-1609
Number of pages13
JournalRNA: a Publication of the RNA Society
Journal publication dateAug 2010
Volume16
Issue8
DOIs
StatePublished

Abstract

The hepatitis C virus internal ribosome entry site (IRES) element contains a three-way junction that is important in the overall RNA conformation, and for its role in the internal initiation of translation. The junction also illustrates some important conformational principles in the folding of three-way helical junctions. It is formally a 3HS(4) junction, with the possibility of two alternative stacking conformers. However, in principle, the junction can also undergo two steps of branch migration that would form 2HS(1)HS(3) and 2HS(2)HS(2) junctions. Comparative gel electrophoresis and ensemble fluorescence resonance energy transfer (FRET) studies show that the junction is induced to fold by the presence of Mg2+ ions in low micromolar concentrations, and suggest that the structure adopted is based on coaxial stacking of the two helices that do not terminate in a hairpin loop (i.e., helix IIId). Single-molecule FRET studies confirm this conclusion, and indicate that there is no minor conformer present based on an alternative choice of helical stacking partners. Moreover, analysis of single-molecule FRET data at an 8-msec resolution failed to reveal evidence for structural transitions. It seems probable that this junction adopts a single conformation as a unique and stable fold.

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