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

191 Citations (Scopus)

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

Fingerprint

Reduced Folate Carrier Protein
Methotrexate
Complementary DNA
Folic Acid
Formyltetrahydrofolates
Cells
Peptides
Human Activities
Sugars
Antineoplastic Agents
Transfection
Clone Cells
Genes
Breast Neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A novel cDNA restores reduced folate carrier activity and methotrexate sensitivity to transport deficient cells. / Dixon, Katharine H.; Lanpher, Brendan C.; Chiu, Jason; Kelley, Kristin; Cowan, Kenneth H.

In: Journal of Biological Chemistry, Vol. 269, No. 1, 07.01.1994, p. 17-20.

Research output: Contribution to journalArticle

Dixon, Katharine H. ; Lanpher, Brendan C. ; Chiu, Jason ; Kelley, Kristin ; Cowan, Kenneth H. / A novel cDNA restores reduced folate carrier activity and methotrexate sensitivity to transport deficient cells. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 1. pp. 17-20.
@article{d5b053e0ae264d60b86d0c72ad4d1b27,
title = "A novel cDNA restores reduced folate carrier activity and methotrexate sensitivity to transport deficient cells",
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.",
author = "Dixon, {Katharine H.} and Lanpher, {Brendan C.} and Jason Chiu and Kristin Kelley and Cowan, {Kenneth H}",
year = "1994",
month = "1",
day = "7",
language = "English (US)",
volume = "269",
pages = "17--20",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "1",

}

TY - JOUR

T1 - A novel cDNA restores reduced folate carrier activity and methotrexate sensitivity to transport deficient cells

AU - Dixon, Katharine H.

AU - Lanpher, Brendan C.

AU - Chiu, Jason

AU - Kelley, Kristin

AU - Cowan, Kenneth H

PY - 1994/1/7

Y1 - 1994/1/7

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0027982786&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027982786&partnerID=8YFLogxK

M3 - Article

VL - 269

SP - 17

EP - 20

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 1

ER -