Human Hepatic glyceraldehyde-3-phosphate Dehydrogenase Binds to the poly(U) Tract of the 3' Non-Coding Region of Hepatitis C Virus Genomic RNA

J Gen Virol. 1999 Dec;80 ( Pt 12):3109-3113. doi: 10.1099/0022-1317-80-12-3109.

Abstract

The unique poly(U/UC) tract, the middle part of the tripartite 3' non-coding region (3'NCR) of hepatitis C virus (HCV) genomic RNA, may represent a recognition signal for the HCV replicase complex. In this study, several proteins binding specifically to immobilized ribooligonucleotide r(U)(25) mimicking this structure were identified using cytosolic extracts from HCV-negative or -positive liver explants, and a prominent 36 kDa protein was studied further. Competition experiments including homoribopolymers revealed binding affinities in the order: oligo/poly(U)>(A)>(C)>(G). The protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a multifunctional protein known to bind RNA. GAPDH bound efficiently to the full-length HCV RNA and binding to various 3'NCR constructs revealed critical dependence upon the presence of the middle part of the 3'NCR. Polypyrimidine tract-binding protein, described previously to bind the 3'NCR, did not bind efficiently to the middle part of 3'NCR and was captured from liver extracts in considerably smaller quantities.

MeSH terms

  • 3' Untranslated Regions / genetics
  • 3' Untranslated Regions / metabolism*
  • Genome, Viral
  • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism*
  • Hepacivirus / genetics*
  • Hepacivirus / metabolism
  • Hepatitis C / virology
  • Humans
  • Liver / enzymology*
  • Liver / virology
  • Oligoribonucleotides / metabolism
  • Poly U / genetics
  • Poly U / metabolism*
  • Polyribonucleotides / metabolism
  • RNA, Viral / genetics
  • RNA, Viral / metabolism*

Substances

  • 3' Untranslated Regions
  • Oligoribonucleotides
  • Polyribonucleotides
  • RNA, Viral
  • Poly U
  • Glyceraldehyde-3-Phosphate Dehydrogenases