Abstract
Recent findings have highlighted the possibility that polymorphisms within the annexin A5 gene (ANXA5) promoter contribute to the etiology of various obstetric complications. However, the underlying mechanisms are unknown. The M2 haplotype of the ANXA5 shows lower activity and less expression of ANXA5 mRNA. This gene promoter region has a motif that potentially forms a G-quadruplex structure. In vitro G-quadruplex propensity estimated by circular dichroism indicated that the M2 haplotype oligonucleotide manifested a decreased potential for G-quadruplex formation. In addition, in vivo G-quadruplex formation of the promoter region was evidenced by the presence of single-stranded DNA shown by sodium bisulfite treatment of placental genomic DNA. Comparative analysis indicated less potential in the M2 allele than the major allele. Promoter activity of the two haplotypes determined by luciferase reporter analysis correlated with the estimated G-quadruplex propensity. Our data lend support to the developing paradigm that genomic variation affects gene expression levels via DNA secondary structures leading to the disease susceptibility.
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Acknowledgements
We thank Ms. Y. Nakura (Osaka Women’s and Children’s Hospital, Japan) for technical assistance, and Drs. H. Kogo, M. Tsutsumi, T. Ohye, M. Shimada (Fujita Health University, Japan), A. Markoff (University of Muenster, Germany), K. Hata (National Research Institute for Child Health and Development, Japan), and K. Shiraki (University of Tsukuba) for helpful discussions. This study was supported by grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (HK and IY), grants-in-aid for Scientific Research from the Ministry of Health, Labor, and Welfare of Japan (HK and IY), and AMED under Grant Number JP17gk0110018 (HK).
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Inagaki, H., Ota, S., Nishizawa, H. et al. Obstetric complication-associated ANXA5 promoter polymorphisms may affect gene expression via DNA secondary structures. J Hum Genet 64, 459–466 (2019). https://doi.org/10.1038/s10038-019-0578-4
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DOI: https://doi.org/10.1038/s10038-019-0578-4
