Mammalian Genome

, Volume 13, Issue 9, pp 483–492

Multiple transcription start sites and alternative splicing in the methylenetetrahydrofolate reductase gene result in two enzyme isoforms

Authors

  • Pamela  Tran
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Daniel  Leclerc
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Manuel  Chan
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Aditya  Pai
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Francois  Hiou-Tim
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Qing  Wu
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Philippe  Goyette
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Carmen  Artigas
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Renate  Milos
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3
  • Rima  Rozen
    • Departments of Human Genetics, Pediatrics, and Biology, McGill University-Montreal Children's Hospital, Montreal, Quebec, Canada H3H 1P3

DOI: 10.1007/s00335-002-2167-6

Cite this article as:
Tran, P., Leclerc, D., Chan, M. et al. Mamm Genome (2002) 13: 483. doi:10.1007/s00335-002-2167-6

Methylenetetrahydrofolate reductase (MTHFR) reduces 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, the major carbon donor in the remethylation of homocysteine to methionine. Mild MTHFR deficiency, due to a common variant at nucleotide 677, has been reported to alter risk for several disorders including cardiovascular disease, neural tube defects, pregnancy complications, and certain cancers. Little is known about MTHFR regulation, since the complete cDNA and gene sequences have not been determined. In earlier work, we isolated and expressed a 2.2-kb human cDNA comprised of 11 coding exons, and we demonstrated that it encoded an active 70-kDa isoform. However, transcript sizes of approximately 7.5 kb and 9.5 kb and the presence of a second isoform of 77 kDa on Western blots suggested that cDNA sequences were incomplete. In this report, we characterized the complete cDNA and gene structure in human and mouse. Variable 5? and 3? UTR regions were identified, resulting in transcript heterogeneity. The 5? and 3? termini of the MTHFR cDNA were found to overlap with the 5? terminus of a chloride ion channel gene (CLCN-6) and the 3? terminus of an unidentified gene, respectively; this finding has resulted in finer mapping of MTHFR on Chromosome (Chr) 1p36.3. Ribonuclease protection assays identified clusters of transcriptional start sites, suggesting the existence of multiple promoters. MTHFR has several polyadenylation sites creating 3?UTR lengths of 0.2 kb–5.0 kb or 0.6 kb–4.0 kb in human and mouse, respectively. In both species, the previously reported exon 1 was redefined to approximately 3.0 kb in length and shown to be alternatively spliced. An important splice variant contains novel coding sequences; this cDNA was expressed and shown to encode the isozyme of 77 kDa. Our results, which suggest intricate regulation of MTHFR, will facilitate additional regulatory and functional studies of the different isoforms.

Copyright information

© Springer-Verlag New York Inc. 2002