Identification of a cis-acting element of human dihydrofolate reductase mRNA

Ningwen Tai, John C. Schmitz, Tian min Chen, Michelle B. O'Neill, Edward Chu

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Human dihydrofolate reductase (DHFR) is a critical target in cancer chemotherapy. Previous studies showed that an 82-nt RNA fragment within the DHFR mRNA protein-coding region functions as a DHFR cis-acting response element. In this study, we further investigated the key elements contained within this sequence that are required for the DHFR mRNA-DHFR protein interaction. Using enzymatic foot-printing assays and RNA-binding experiments, we isolated a 27-nt sequence (DHFR27, corresponding to nts 407-433), which bound with high affinity and specificity to human DHFR to form a ribonucleoprotein complex. In vivo transient transfection experiments using a luciferase reporter system revealed that DHFR27 RNA could repress the luciferase expression in a DHFR-dependent manner when placed upstream of luciferase mRNA. This work provides new insights into the essential molecular elements that mediate RNA-protein interactions.

Original languageEnglish (US)
Pages (from-to)795-800
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume369
Issue number3
DOIs
StatePublished - May 9 2008
Externally publishedYes

Keywords

  • Dihydrofolate reductase
  • RNA-protein interaction
  • Translational regulation
  • cis-acting element

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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