Specific Binding of Human Dihydrofolate Reductase Protein to Dihydrofolate Reductase Messenger RNA in Vitro

Edward Chu, Chris H. Takimoto, Donna Voeller, Jean L. Grem, Carmen J. Allegra

Research output: Contribution to journalArticle

113 Scopus citations

Abstract

Dihydrofolate reductase (DHFR) is a critical enzyme in de novo purine and thymidylate biosynthesis. An RNA gel mobility shift assay was used to demonstrate a specific interaction between human recombinant DHFR protein and its corresponding DHFR mRNA. Incubation of DHFR protein with either its substrates, dihydrofolate or NADPH, or with an inhibitor, methotrexate, repressed its ability to interact with DHFR mRNA. An in vitro rabbit reticulocyte lysate translation system was used to show that the addition of exogenous human recombinant DHFR protein to in vitro translation reactions specifically inhibited DHFR mRNA translation. These studies suggest that the direct interaction between DHFR protein and its mRNA may be a mechanism for regulation of DHFR synthesis.

Original languageEnglish (US)
Pages (from-to)4756-4760
Number of pages5
JournalBiochemistry
Volume32
Issue number18
DOIs
StatePublished - May 1 1993

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ASJC Scopus subject areas

  • Biochemistry

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