Secondary structure of the Irf7 5'-UTR, analyzed using SHAPE (selective 2'-hydroxyl acylation analyzed by primer extension)

BMB Rep. 2014 Oct;47(10):558-62. doi: 10.5483/bmbrep.2014.47.10.281.

Abstract

OASL1 is a member of the 2'-5'-oligoadenylate synthetase (OAS) family and promotes viral clearance by activating RNase L. OASL1 interacts with the 5'-untranslated region (UTR) of interferon regulatory factor 7 (Irf7) and inhibits its translation. To identify the secondary structure required for OASL1 binding, we examined the 5'-UTR of the Irf7 transcript using "selective 2'-hydroxyl acylation analyzed by primer extension" (SHAPE). SHAPE takes advantage of the selective acylation of residues in single-stranded regions by 1-methyl-7-nitroisatoic anhydride (1M7). We found five major acylation sites located in, or next to, predicted single-stranded regions of the Irf7 5'-UTR. These results demonstrate the involvement of the stem structure of the Irf7 5'-UTR in the regulation of Irf7 translation, mediated by OASL1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions / genetics*
  • Acylation
  • Adenosine / metabolism
  • Base Sequence
  • DNA Primers / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Hydroxylation
  • Interferon Regulatory Factor-7 / genetics*
  • Molecular Sequence Data
  • Nucleic Acid Conformation*
  • Oxazines / metabolism
  • Polymerase Chain Reaction / methods*

Substances

  • 1-methyl-7-nitroisatoic anhydride
  • 5' Untranslated Regions
  • DNA Primers
  • Interferon Regulatory Factor-7
  • Oxazines
  • Adenosine