A novel cDNA restores reduced folate carrier activity and methotrexate sensitivity to transport deficient cells

Katharine H. Dixon, Brendan C. Lanpher, Jason Chiu, Kristin Kelley, Kenneth H. Cowan

Research output: Contribution to journalArticle

194 Scopus citations

Abstract

Mammalian cells accumulate reduced folates and methotrexate, a folate antagonist, through the reduced-folate carrier (RFC) (Goldman, I. D., Lichtenstein, N. S., and Oliverio, V. T. (1968) J. Biol. Chem. 243, 5007- 5017). This study describes the isolation and expression of a cDNA clone that restores RFC activity to human breast cancer cells defective in this transporter. The cDNA codes for a peptide (mRFC1) of 58 kDa, whose hydropathy plot, resembling those of mammalian sugar transporters, predicts that it may be a member of a superfamily of transporter genes. Transfection of methotrexate-resistant (MTX(R)) ZR-75-1 cells with an expression vector, pRFC1, that codes for this peptide restores their ability to accumulate methotrexate. Furthermore, transport of methotrexate into pRFC1-transfected cells is blocked by a 10-fold molar excess of the reduced folate, 5- formyltetrahydrofolic acid, but is unaffected by folic acid. The increase in methotrexate uptake that is observed in pRFC1-transfected MTX(R) ZR-75-1 cells reverses their resistance to this antitumor agent.

Original languageEnglish (US)
Pages (from-to)17-20
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number1
StatePublished - Jan 7 1994

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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