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Aberrant mRNA splicing of paired box 4 (PAX4) IVS7-1G>A mutation causing maturity-onset diabetes of the young, type 9

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Abstract

Aims

Paired box 4 (PAX4) mutations cause maturity-onset diabetes of the young, type 9 (MODY9). The molecular defect and alteration of PAX4 function associated with the mutation PAX4 IVS7-1G>A in a family with MODY9 and severe diabetic complications were studied.

Methods

We investigated the functional consequences of PAX4 IVS7-1G>A on mRNA splicing using minigene assays. Wild-type and mutant PAX4 were expressed in mouse pancreatic β- and α-cell lines, and protein levels and translocation of PAX4 into the nucleus were determined. We also examined transcriptional repression of PAX4 target-gene promoters and β-cell viability under diabetic-like (high-glucose) conditions.

Results

PAX4 IVS7-1G>A disrupts an acceptor splice site, causing an adjacent cryptic splice site within exon 8 to be used, resulting in a three-nucleotide deletion and glutamine deletion at position 250 (p.Q250del). Wild-type and PAX4 Q250del proteins were expressed at similar levels and could translocate normally into the nucleus in βTC3 and αTC1.9 cells. However, the repressor functions of PAX4 Q250del on human insulin and glucagon promoters in INS-1 832/13 and αTC1.9 cells were significantly decreased, compared with that of wild-type PAX4. Moreover, the rate of apoptosis was increased in INS-1 cells over-expressing PAX4 Q250del when cultured in high-glucose conditions.

Conclusions

PAX4 IVS7-1G>A caused aberrant mRNA splicing and PAX4 Q250 deletion. The mutation impaired PAX4 repressor functions on target-gene promoters and increased susceptibility to apoptosis upon high glucose exposure. Thus, PAX4 IVS7-1G>A contributes to the pathogenesis of diabetes in this MODY9 family through β-cell dysfunction.

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Acknowledgments

JS was supported by the joint Royal Golden Jubilee (RGJ) PhD Program of Thailand Research Fund (TRF) and Mahidol University (Grant No. PHD/0096/2550). This work was supported by a Siriraj Research Development Grant, a Faculty of Medicine Siriraj Hospital, Mahidol University Grant (to SK and NP). NP was supported by a TRF Research Career Development Grant and the Office of the Higher Education Commission and Mahidol University under the National Research Universities Initiative. PY was supported by a Siriraj Charoemprakiat Grant and a TRF Senior Research Scholar Award. JS conducted experiments, analyzed data, and prepared the manuscript. SK, NC, and WT conducted experiments and analyzed data. NP corresponded with patients, collected clinical data, discussed the experimental results, and edited the manuscript. PY planned the project and experiments, discussed the results, and edited the manuscript.

Conflict of interest

The authors declare no conflicts of interest.

Ethical standard

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Siriraj Institutional Review Board, Bangkok, Thailand, and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Human and animal rights

This article does not contain any studies with animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants for being included in the study.

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Authors and Affiliations

  1. Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Jatuporn Sujjitjoon & Pa-thai Yenchitsomanus

  2. Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Jatuporn Sujjitjoon

  3. Department of Physiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Suwattanee Kooptiwut

  4. Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Nalinee Chongjaroen & Watip Tangjittipokin

  5. Division of Endocrinology and Metabolism, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Nattachet Plengvidhya

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  1. Jatuporn Sujjitjoon
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  2. Suwattanee Kooptiwut
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  5. Nattachet Plengvidhya
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Correspondence to Nattachet Plengvidhya or Pa-thai Yenchitsomanus.

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Sujjitjoon, J., Kooptiwut, S., Chongjaroen, N. et al. Aberrant mRNA splicing of paired box 4 (PAX4) IVS7-1G>A mutation causing maturity-onset diabetes of the young, type 9. Acta Diabetol 53, 205–216 (2016). https://doi.org/10.1007/s00592-015-0760-x

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  • DOI: https://doi.org/10.1007/s00592-015-0760-x

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