Organization of the murine reduced folate carrier gene and identification of variant splice forms

Kevin E. Brigle, Michael J. Spinella, Esteban E. Sierra, I. David Goldman

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

RT-PCR analysis of the reduced folate carrier (RFC) from L1210 and murine erythroleukemia cells led to the identification of three clones which appeared to result from the use of alternative splice sites. The nucleotide sequence of each splice form predicts a protein that contains at least the first 7 transmembrane domains of the parental RFC protein followed by a novel hydrophilic carboxyl terminus of 33, 72, or 105 amino acid residues. Sequence analysis of cDNA clones isolated from murine liver and the results of 5'- RACE from L1210 cells indicated that RFC also utilizes alternate 5'-terminal exons. To understand how the alternatively spliced RFC transcripts and multiple 5'-termini were generated, the genomic organization of RFC was determined. The gene is comprised of at least 8 exons, the first two of which encode the alternative 5' termini. Based on sequence identity with cDNAs encoding RFC from hamster and rat, however, it appears that additional 5' exons may be present. Two of the RFC splice variants result from the use of a cryptic splice donor site within exon 4 and the third results from the use of a cryptic splice acceptor site within exon 5. In addition, the splice variant form that encodes the largest protein also utilizes an alternative exon located between exons 5 and 6. The apparent use of alternative transcriptional start sites and the identification of several RFC splice forms raises the possibility that unique RFC molecules may be generated that exhibit tissue- or cell line-specific distribution.

Original languageEnglish (US)
Pages (from-to)191-198
Number of pages8
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1353
Issue number2
DOIs
StatePublished - Aug 7 1997

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Reduced Folate Carrier Protein
RNA Splice Sites
Genes
Exons
Complementary DNA
Clone Cells
Leukemia, Erythroblastic, Acute
Cricetinae
Sequence Analysis
Liver
Proteins
Rats
Nucleotides
Cells
Amino Acids
Cell Line
Polymerase Chain Reaction

Keywords

  • Folate transport
  • Genomic organization
  • Methotrexate transport
  • Reduced folate carrier
  • Reduced folate carrier 1
  • Splice form

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology
  • Biophysics

Cite this

Organization of the murine reduced folate carrier gene and identification of variant splice forms. / Brigle, Kevin E.; Spinella, Michael J.; Sierra, Esteban E.; Goldman, I. David.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1353, No. 2, 07.08.1997, p. 191-198.

Research output: Contribution to journalArticle

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