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Analysis of the MTHFD1 promoter and risk of neural tube defects

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Abstract

Genetic variants in MTHFD1 (5,10-methylenetetrahydrofolate dehydrogenase/5,10-methenyltetrahydrofolate cyclohydrolase/ 10-formyltetrahydrofolate synthetase), an important folate metabolic enzyme, are associated with a number of common diseases, including neural tube defects (NTDs). This study investigates the promoter of the human MTHFD1 gene in a bid to understand how this gene is controlled and regulated. Following a combination of in silico and molecular approaches, we report that MTHFD1 expression is controlled by a TATA-less, Initiator-less promoter and transcription is initiated at multiple start sites over a 126 bp region. We confirmed the presence of three database polymorphisms (dbSNP) by direct sequencing of the upstream region (rs1076991 C > T, rs8010584 G > A, rs4243628 G > T), with a fourth (dbSNP rs746488 A > T) not found to be polymorphic in our population and no novel polymorphisms identified. We demonstrate that a common SNP rs1076991 C > T within the window of transcriptional initiation exerts a significant effect on promoter activity in vitro. We investigated this SNP as a potential risk factor for NTDs in a large homogenous Irish population and determined that it is not an independent risk factor, but, it does increase both case (χ 2 = 11.06, P = 0.001) and maternal (χ 2 = 6.68, P = 0.01) risk when allele frequencies were analysed in combination with the previously identified disease-associated p.R653Q (c.1958 G > A; dbSNP rs2236225) polymorphism. These results provide the first insight into how MTHFD1 is regulated and further emphasise its importance during embryonic development.

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Acknowledgments

We wish to thank the Irish Association for Spina Bifida and Hydrocephalus and the Public Health Nurses for assistance with subject recruitment and the following Dublin maternity hospitals for control recruitment: National Maternity Hospital Holles street, the Coombe Womens’ Hospital and the Rotunda Hospital. This research was supported by the Intramural Research Program of the NICHD, NIH and the Health Research Board of Ireland. Approval was obtained from the Research Ethics Committee of the Health Research Board, Ireland and the Institutional Review Board at the National Institutes of Health. All research participants provided informed consent.

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Authors and Affiliations

  1. Nutritional Genomics Group, School of Biotechnology, Dublin City University, Dublin 9, Ireland

    Nicola Carroll & Anne Parle-McDermott

  2. Molecular Pathogenesis Section, Genome Technology Branch, National Human Genome Research Institute, Building 50, Room 5306, 50 South Drive, MSC 8004, Bethesda, MD, 20892-8004, USA

    Faith Pangilinan & Lawrence C. Brody

  3. School of Medicine, Trinity College Dublin, Dublin 2, Ireland

    Anne M. Molloy

  4. Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, USA

    James Troendle & James L. Mills

  5. Child Health Epidemiology Division, Health Research Board, Dublin, Ireland

    Peadar N. Kirke

  6. School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland

    John M. Scott

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  1. Nicola Carroll
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  2. Faith Pangilinan
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  3. Anne M. Molloy
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  4. James Troendle
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  9. Anne Parle-McDermott
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Correspondence to Anne Parle-McDermott.

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Carroll, N., Pangilinan, F., Molloy, A.M. et al. Analysis of the MTHFD1 promoter and risk of neural tube defects. Hum Genet 125, 247–256 (2009). https://doi.org/10.1007/s00439-008-0616-3

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  • DOI: https://doi.org/10.1007/s00439-008-0616-3

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