Molecular basis for decreased folylpoly-γ-glutamate synthetase expression in a methotrexate resistant CCRF-CEM mutant cell line

Guy J. Leclerc, Teresa A. York, Tingting Hsieh-Kinser, Julio Barredo

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A CCRF-CEM mutant, CEM-p, has been shown to exhibit resistance to methotrexate due to decreased methotrexate polyglutamate accumulation. To ascertain the mechanism(s) responsible for this phenotype, we analyzed FPGS and SLC19A1 mRNA expression, examined FPGS promoter activity, and determined nucleotide sequence of the FPGS promoter and full length cDNA from CCRF-CEM and CEM-p cells. We identified in FPGS from CEM-p cells three amino acid substitutions that altered the ATP binding P-loop, glutamate/folate binding, and a conserved domain located at the carboxyl-terminal. Our data demonstrated for the first time the importance of the highly conserved domain (VTGSLHLVGGV) located at the carboxyl-terminal for FPGS activity.

Original languageEnglish
Pages (from-to)293-299
Number of pages7
JournalLeukemia Research
Volume31
Issue number3
DOIs
StatePublished - Mar 1 2007
Externally publishedYes

Fingerprint

Ligases
Methotrexate
Glutamic Acid
Cell Line
Amino Acid Substitution
Folic Acid
Complementary DNA
Adenosine Triphosphate
Phenotype
Messenger RNA
methotrexate polyglutamate

Keywords

  • CCRF-CEM
  • Folate
  • FPGS
  • Leukemia
  • Methotrexate resistance
  • Mutagenesis
  • Mutant
  • Point mutation
  • SLC19A1
  • T-ALL

ASJC Scopus subject areas

  • Cancer Research
  • Hematology
  • Oncology

Cite this

Molecular basis for decreased folylpoly-γ-glutamate synthetase expression in a methotrexate resistant CCRF-CEM mutant cell line. / Leclerc, Guy J.; York, Teresa A.; Hsieh-Kinser, Tingting; Barredo, Julio.

In: Leukemia Research, Vol. 31, No. 3, 01.03.2007, p. 293-299.

Research output: Contribution to journalArticle

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