nach oben

08.05.2015 | Original Article

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

Erschienen in: Acta Diabetologica | Ausgabe 2/2016

Einloggen, um Zugang zu erhalten

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.

Fajans SS, Bell GI, Polonsky KS (2001) Molecular mechanisms and clinical pathophysiology of maturity-onset diabetes of the young. N Engl J Med 345(13):971–980CrossRefPubMed

Yamagata K, Furuta H, Oda N, Kaisaki PJ, Menzel S, Cox NJ et al (1996) Mutations in the hepatocyte nuclear factor-4alpha gene in maturity-onset diabetes of the young (MODY1). Nature 384(6608):458–460CrossRefPubMed

Froguel P (1996) Glucokinase and MODY: from the gene to the disease. Diabet Med 13(9 Suppl 6):S96–S97PubMed

Yamagata K, Oda N, Kaisaki PJ, Menzel S, Furuta H, Vaxillaire M et al (1996) Mutations in the hepatocyte nuclear factor-1 alpha gene in maturity-onset diabetes of the young (MODY3). Nature 384(6608):455–458CrossRefPubMed

Stoffers DA, Ferrer J, Clarke WL, Habener JF (1997) Early-onset type-II diabetes mellitus (MODY4) linked to IPF1. Nat Genet 17(2):138–139CrossRefPubMed

Horikawa Y, Iwasaki N, Hara M, Furuta H, Hinokio Y, Cockburn BN et al (1997) Mutation in hepatocyte nuclear factor-1 beta gene (TCF2) associated with MODY. Nat Genet 17(4):384–385CrossRefPubMed

Malecki MT, Jhala US, Antonellis A, Fields L, Doria A, Orban T et al (1999) Mutations in NEUROD1 are associated with the development of type 2 diabetes mellitus. Nat Genet 23(3):323–328CrossRefPubMed

Bell GI, Froguel P, Nishi S, Pilkis SJ, Stoffel M, Takeda J et al (1993) Mutations of the human glucokinase gene and diabetes mellitus. Trends Endocrinol Metab 4(3):86–90CrossRefPubMed

Plengvidhya N, Boonyasrisawat W, Chongjaroen N, Jungtrakoon P, Sriussadaporn S, Vannaseang S et al (2009) Mutations of maturity-onset diabetes of the young (MODY) genes in Thais with early-onset type 2 diabetes mellitus. Clin Endocrinol 70(6):847–853CrossRef

10.

Lee HJ, Ahn CW, Kim SJ, Song YD, Lim SK, Kim KR et al (2001) Mutation in hepatocyte nuclear factor-1alpha is not a common cause of MODY and early-onset type 2 diabetes in Korea. Acta Diabetol 38(3):123–127CrossRefPubMed

11.

Chapla A, Mruthyunjaya MD, Asha HS, Varghese D, Varshney M, Vasan SK et al (2015) Maturity-onset diabetes of the young in India—a distinctive mutation pattern identified through targeted next-generation sequencing. Clin Endocrinol 82(4):533–542CrossRef

12.

Plengvidhya N, Kooptiwut S, Songtawee N, Doi A, Furuta H, Nishi M et al (2007) PAX4 mutations in Thais with maturity onset diabetes of the young. J Clin Endocrinol Metab 92(7):2821–2826CrossRefPubMed

13.

Kooptiwut S, Plengvidhya N, Chukijrungroat T, Sujjitjoon J, Semprasert N, Furuta H et al (2012) Defective PAX4 R192H transcriptional repressor activities associated with maturity onset diabetes of the young and early onset-age of type 2 diabetes. J Diabetes Complicat 26(4):343–347CrossRefPubMed

14.

Biason-Lauber A, Boehm B, Lang-Muritano M, Gauthier BR, Brun T, Wollheim CB et al (2005) Association of childhood type 1 diabetes mellitus with a variant of PAX4: possible link to beta cell regenerative capacity. Diabetologia 48(5):900–905CrossRefPubMed

15.

Shimajiri Y, Sanke T, Furuta H, Hanabusa T, Nakagawa T, Fujitani Y et al (2001) A missense mutation of Pax4 gene (R121W) is associated with type 2 diabetes in Japanese. Diabetes 50(12):2864–2869CrossRefPubMed

16.

Shimajiri Y, Shimabukuro M, Tomoyose T, Yogi H, Komiya I, Takasu N (2003) PAX4 mutation (R121W) as a prodiabetic variant in Okinawans. Biochem Biophys Res Commun 302(2):342–344CrossRefPubMed

17.

Mauvais-Jarvis F, Smith SB, Le May C, Leal SM, Gautier JF, Molokhia M, Riveline JP, Rajan AS, Kevorkian JP, Zhang S, Vexiau P, German MS, Vaisse C (2004) PAX4 gene variations predispose to ketosis-prone diabetes. Hum Mol Gen 13(24):3151–3159CrossRefPubMed

18.

Sosa-Pineda B (2004) The gene Pax4 is an essential regulator of pancreatic beta-cell development. Mol Cells 18(3):289–294PubMed

19.

Sosa-Pineda B, Chowdhury K, Torres M, Oliver G, Gruss P (1997) The Pax4 gene is essential for differentiation of insulin-producing beta cells in the mammalian pancreas. Nature 386(6623):399–402CrossRefPubMed

20.

Brun T, Franklin I, St-Onge L, Biason-Lauber A, Schoenle EJ, Wollheim CB et al (2004) The diabetes-linked transcription factor PAX4 promotes beta-cell proliferation and survival in rat and human islets. J Cell Biol 167(6):1123–1135CrossRefPubMedPubMedCentral

21.

Smith SB, Ee HC, Conners JR, German MS (1999) Paired-homeodomain transcription factor PAX4 acts as a transcriptional repressor in early pancreatic development. Mol Cell Biol 19(12):8272–8280CrossRefPubMedPubMedCentral

22.

He KH, Lorenzo PI, Brun T, Jimenez Moreno CM, Aeberhard D, Ortega JV et al (2011) In vivo conditional Pax4 overexpression in mature islet beta-cells prevents stress-induced hyperglycemia in mice. Diabetes 60(6):1705–1715CrossRefPubMedCentral

23.

Jo W, Endo M, Ishizu K, Nakamura A, Tajima T (2011) A novel PAX4 mutation in a Japanese patient with maturity-onset diabetes of the young. Tohoku J Exp Med 223(2):113–118CrossRefPubMed

24.

Gaildrat P, Killian A, Martins A, Tournier I, Frebourg T, Tosi M (2010) Use of splicing reporter minigene assay to evaluate the effect on splicing of unclassified genetic variants. Methods Mol Biol 653:249–257CrossRefPubMed

25.

Cooper TA (2005) Use of minigene systems to dissect alternative splicing elements. Methods 37(4):331–340CrossRefPubMed

26.

Petersen HV, Jorgensen MC, Andersen FG, Jensen J, Tove F, Jorgensen R et al (2000) Pax4 represses pancreatic glucagon gene expression. Mol Cell Biol Res Commun 3(4):249–254CrossRefPubMed

27.

Gamez-Pozo A, Palacios I, Kontic M, Menendez I, Camino I, Garcia-Miguel P, Abelairas J, Pestana A, Alonso J (2007) Pathogenic validation of unique germline intronic variants of RB1 in retinoblastoma patients using minigenes. Hum Mutat 28(12):1245CrossRefPubMed

28.

Weisschuh N, Wissinger B, Gramer E (2012) A splice site mutation in the PAX6 gene which induces exon skipping causes autosomal dominant inherited aniridia. Mol Vis 18:751–757PubMedPubMedCentral

29.

Nagamura Y, Yamazaki M, Shimazu S, Sano K, Tsukada T, Sakurai A (2012) A novel splice site mutation of the MEN1 gene identified in a patient with primary hyperparathyroidism. Endocr J 59(6):523–530CrossRefPubMed

30.

Brun T, He KH, Lupi R, Boehm B, Wojtusciszyn A, Sauter N et al (2008) The diabetes-linked transcription factor Pax4 is expressed in human pancreatic islets and is activated by mitogens and GLP-1. Hum Mol Genet 17(4):478–489CrossRefPubMed

31.

Weir GC, Sharma A, Zangen DH, Bonner-Weir S (1997) Transcription factor abnormalities as a cause of beta cell dysfunction in diabetes: a hypothesis. Acta Diabetol 34(3):177–184CrossRefPubMed

32.

Rath MF, Bailey MJ, Kim JS, Coon SL, Klein DC, Moller M (2009) Developmental and daily expression of the Pax4 and Pax6 homeobox genes in the rat retina: localization of Pax4 in photoreceptor cells. J Neurochem 108(1):285–294CrossRefPubMed

33.

Reichman S, Kalathur RK, Lambard S, Ait-Ali N, Yang Y, Lardenois A et al (2010) The homeobox gene CHX10/VSX2 regulates RdCVF promoter activity in the inner retina. Hum Mol Genet 19(2):250–261CrossRefPubMedPubMedCentral

34.

Jacobson PA, Schladt D, Israni A, Oetting WS, Lin YC, Leduc R et al (2012) Genetic and clinical determinants of early, acute calcineurin inhibitor-related nephrotoxicity: results from a kidney transplant consortium. Transplantation 93(6):624–631PubMedPubMedCentral

Titel: Aberrant mRNA splicing of paired box 4 (PAX4) IVS7-1G>A mutation causing maturity-onset diabetes of the young, type 9
Publikationsdatum: 08.05.2015
Erschienen in: Acta Diabetologica / Ausgabe 2/2016
Print ISSN: 0940-5429
Elektronische ISSN: 1432-5233
DOI: https://doi.org/10.1007/s00592-015-0760-x

Leitlinien kompakt für die Innere Medizin

Mit medbee Pocketcards sicher entscheiden.

^{Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag}

Kostenlos registrieren

Neu im Fachgebiet Innere Medizin

Notfall-TEP der Hüfte ist auch bei 90-Jährigen machbar

26.04.2024 Hüft-TEP Nachrichten

Ob bei einer Notfalloperation nach Schenkelhalsfraktur eine Hemiarthroplastik oder eine totale Endoprothese (TEP) eingebaut wird, sollte nicht allein vom Alter der Patientinnen und Patienten abhängen. Auch über 90-Jährige können von der TEP profitieren.

Niedriger diastolischer Blutdruck erhöht Risiko für schwere kardiovaskuläre Komplikationen

25.04.2024 Hypotonie Nachrichten

Wenn unter einer medikamentösen Hochdrucktherapie der diastolische Blutdruck in den Keller geht, steigt das Risiko für schwere kardiovaskuläre Ereignisse: Darauf deutet eine Sekundäranalyse der SPRINT-Studie hin.

Bei schweren Reaktionen auf Insektenstiche empfiehlt sich eine spezifische Immuntherapie

25.04.2024 Allergien und Intoleranzreaktionen Nachrichten

Insektenstiche sind bei Erwachsenen die häufigsten Auslöser einer Anaphylaxie. Einen wirksamen Schutz vor schweren anaphylaktischen Reaktionen bietet die allergenspezifische Immuntherapie. Jedoch kommt sie noch viel zu selten zum Einsatz.

Therapiestart mit Blutdrucksenkern erhöht Frakturrisiko

25.04.2024 Hypertonie Nachrichten

Beginnen ältere Männer im Pflegeheim eine Antihypertensiva-Therapie, dann ist die Frakturrate in den folgenden 30 Tagen mehr als verdoppelt. Besonders häufig stürzen Demenzkranke und Männer, die erstmals Blutdrucksenker nehmen. Dafür spricht eine Analyse unter US-Veteranen.

Update Innere Medizin

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.

Newsletter bestellen