Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Diabetologia
Published in: Diabetologia 6/2013

01-06-2013 | Article

Multiple functional polymorphisms in the G6PC2 gene contribute to the association with higher fasting plasma glucose levels

Authors: D. A. Baerenwald, A. Bonnefond, N. Bouatia-Naji, B. P. Flemming, O. C. Umunakwe, J. K. Oeser, L. D. Pound, N. L. Conley, S. Cauchi, S. Lobbens, E. Eury, B. Balkau, O. Lantieri, P. K. Dadi, D. A. Jacobson, P. Froguel, R. M. O’Brien, MAGIC Investigators

Published in: Diabetologia | Issue 6/2013

Login to get access

Abstract

Aims/hypothesis

We previously identified the G6PC2 locus as a strong determinant of fasting plasma glucose (FPG) and showed that a common G6PC2 intronic single nucleotide polymorphism (SNP) (rs560887) and two common G6PC2 promoter SNPs (rs573225 and rs13431652) are highly associated with FPG. However, these promoter SNPs have complex effects on G6PC2 fusion gene expression, and our data suggested that only rs13431652 is a potentially causative SNP. Here we examine the effect of rs560887 on G6PC2 pre-mRNA splicing and the contribution of an additional common G6PC2 promoter SNP, rs2232316, to the association signal.

Methods

Minigene analyses were used to characterise the effect of rs560887 on G6PC2 pre-mRNA splicing. Fusion gene and gel retardation analyses characterised the effect of rs2232316 on G6PC2 promoter activity and transcription factor binding. The genetic association of rs2232316 with FPG variation was assessed using regression adjusted for age, sex and BMI in 4,220 Europeans with normal FPG.

Results

The rs560887-G allele was shown to enhance G6PC2 pre-mRNA splicing, whereas the rs2232316-A allele enhanced G6PC2 transcription by promoting Foxa2 binding. Genetic analyses provide evidence for association of the rs2232316-A allele with increased FPG (β = 0.04 mmol/l; p = 4.3 × 10−3) as part of the same signal as rs560887, rs573225 and rs13431652.

Conclusions/interpretation

As with rs13431652, the in situ functional data with rs560887 and rs2232316 are in accord with the putative function of G6PC2 in pancreatic islets, and suggest that all three are potentially causative SNPs that contribute to the association between G6PC2 and FPG.
Appendix
Available only for authorised users
Literature
1.
Hutton JC, O’Brien RM (2009) The glucose-6-phosphatase catalytic subunit gene family. J Biol Chem 284:29241–29245PubMedCrossRef
2.
Arden SD, Zahn T, Steegers S et al (1999) Molecular cloning of a pancreatic islet-specific glucose-6-phosphatase catalytic subunit-related protein. Diabetes 48:531–542PubMedCrossRef
3.
Martin CC, Bischof LJ, Bergman B et al (2001) Cloning and characterization of the human and rat islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) genes. J Biol Chem 276:25197–25207PubMedCrossRef
4.
Wang Y, Martin CC, Oeser JK et al (2007) Deletion of the gene encoding the islet-specific glucose-6-phosphatase catalytic subunit-related protein autoantigen results in a mild metabolic phenotype. Diabetologia 50:774–778PubMedCrossRef
5.
Hutton JC, Eisenbarth GS (2003) A pancreatic beta-cell-specific homolog of glucose-6-phosphatase emerges as a major target of cell-mediated autoimmunity in diabetes. Proc Natl Acad Sci U S A 100:8626–8628PubMedCrossRef
6.
Bouatia-Naji N, Rocheleau G, van Lommel L et al (2008) A polymorphism within the G6PC2 gene is associated with fasting plasma glucose levels. Science 320:1085–1088PubMedCrossRef
7.
Chen WM, Erdos MR, Jackson AU et al (2008) Variations in the G6PC2/ABCB11 genomic region are associated with fasting glucose levels. J Clin Invest 118:2620–2628PubMed
8.
Droumaguet C, Balkau B, Simon D et al (2006) Use of HbA1c in predicting progression to diabetes in French men and women: data from an Epidemiological Study on the Insulin Resistance Syndrome (DESIR). Diabetes Care 29:1619–1625PubMedCrossRef
9.
Abdul-Ghani MA, DeFronzo RA (2009) Plasma glucose concentration and prediction of future risk of type 2 diabetes. Diabetes Care 32(Suppl 2):S194–S198PubMedCrossRef
10.
Edelman D, Olsen MK, Dudley TK, Harris AC, Oddone EZ (2004) Utility of hemoglobin A1c in predicting diabetes risk. J Gen Intern Med 19:1175–1180PubMedCrossRef
11.
Coutinho M, Gerstein HC, Wang Y, Yusuf S (1999) The relationship between glucose and incident cardiovascular events. A metaregression analysis of published data from 20 studies of 95,783 individuals followed for 12.4 years. Diabetes Care 22:233–240PubMedCrossRef
12.
Khaw KT, Wareham N, Luben R et al (2001) Glycated haemoglobin, diabetes, and mortality in men in Norfolk cohort of European prospective investigation of cancer and nutrition (EPIC-Norfolk). BMJ 322:15–18PubMedCrossRef
13.
Sarwar N, Gao P, Seshasai SR et al (2010) Diabetes mellitus, fasting blood glucose concentration, and risk of vascular disease: a collaborative meta-analysis of 102 prospective studies. Lancet 375:2215–2222PubMedCrossRef
14.
Prokopenko I, Langenberg C, Florez JC et al (2008) Variants in MTNR1B influence fasting glucose levels. Nat Genet 41:77–81PubMedCrossRef
15.
Bouatia-Naji N, Bonnefond A, Cavalcanti-Proenca C et al (2009) A variant near MTNR1B is associated with increased fasting plasma glucose levels and type 2 diabetes risk. Nat Genet 41:89–94PubMedCrossRef
16.
Reiling E, van ’t Riet E, Groenewoud MJ et al (2009) Combined effects of single-nucleotide polymorphisms in GCK, GCKR, G6PC2 and MTNR1B on fasting plasma glucose and type 2 diabetes risk. Diabetologia 52:1866–1870PubMedCrossRef
17.
Dupuis J, Langenberg C, Prokopenko I et al (2010) New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk. Nat Genet 42:105–116PubMedCrossRef
18.
Hu C, Zhang R, Wang C et al (2009) A genetic variant of G6PC2 is associated with type 2 diabetes and fasting plasma glucose level in the Chinese population. Diabetologia 52:451–456PubMedCrossRef
19.
Newgard CB, Lu D, Jensen MV et al (2002) Stimulus/secretion coupling factors in glucose-stimulated insulin secretion: insights gained from a multidisciplinary approach. Diabetes 51(Suppl 3):S389–S393PubMedCrossRef
20.
Bouatia-Naji N, Bonnefond A, Baerenwald DA et al (2010) Genetic and functional assessment of the role of the rs13431652-A and rs573225-A alleles in the G6PC2 promoter that strongly associate with elevated fasting glucose levels. Diabetes 59:2662–2671PubMedCrossRef
21.
Voight BF, Kang HM, Ding J et al (2012) The Metabochip, a custom genotyping array for genetic studies of metabolic, cardiovascular, and anthropometric traits. PLoS Genetics 8:e1002793PubMedCrossRef
22.
Singh G, Cooper TA (2006) Minigene reporter for identification and analysis of cis elements and trans factors affecting pre-mRNA splicing. Biotechniques 41:177–181PubMedCrossRef
23.
Cooper TA (1998) Muscle-specific splicing of a heterologous exon mediated by a single muscle-specific splicing enhancer from the cardiac troponin T gene. Mol Cell Biol 18:4519–4525PubMed
24.
Martin CC, Oeser JK, O’Brien RM (2004) Differential regulation of islet-specific glucose-6-phosphatase catalytic subunit-related protein gene transcription by Pax-6 and Pdx-1. J Biol Chem 279:34277–34289PubMedCrossRef
25.
Martin C, Flemming B, Wang Y, Oeser J, O’Brien R (2008) Foxa2 and MafA regulate islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP/G6PC2) gene expression. J Mol Endocrinol 41:315–328PubMedCrossRef
26.
Dogra RS, Vaidyanathan P, Prabakar KR, Marshall KE, Hutton JC, Pugliese A (2006) Alternative splicing of G6PC2, the gene coding for the islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP), results in differential expression in human thymus and spleen compared with pancreas. Diabetologia 49:953–957PubMedCrossRef
27.
Stoss O, Stoilov P, Hartmann AM, Nayler O, Stamm S (1999) The in vivo minigene approach to analyze tissue-specific splicing. Brain Res Brain Res Protoc 4:383–394PubMedCrossRef
28.
Cooper TA (2005) Use of minigene systems to dissect alternative splicing elements. Methods 37:331–340PubMedCrossRef
29.
Ebert DH, Bischof LJ, Streeper RS et al (1999) Structure and promoter activity of an islet-specific glucose-6-phosphatase catalytic subunit-related gene. Diabetes 48:543–551PubMedCrossRef
30.
Jackson IJ (1991) A reappraisal of non-consensus mRNA splice sites. Nucleic Acids Res 19:3795–3798PubMedCrossRef
31.
Bischof LJ, Martin CC, Svitek CA et al (2001) Characterization of the mouse islet-specific glucose-6-phosphatase catalytic subunit-related protein gene promoter by in situ footprinting. Correlation with fusion gene expression in the islet derived βTC-3 and hamster insulinoma tumor cell lines. Diabetes 50:502–514PubMedCrossRef
32.
Frigeri C, Martin CC, Svitek CA et al (2004) The proximal islet-specific glucose-6-phosphatase catalytic subunit related protein (IGRP) autoantigen promoter is sufficient to initiate but not maintain transgene expression in mouse islets in vivo. Diabetes 53:1754–1764PubMedCrossRef
33.
Martin CC, Oeser JK, Svitek CA, Hunter SI, Hutton JC, O’Brien RM (2002) Identification and characterization of a human cDNA and gene encoding a ubiquitously expressed glucose-6-phosphatase catalytic subunit-related protein. J Mol Endocrinol 29:205–222PubMedCrossRef
34.
Quandt K, Frech K, Karas H, Wingender E, Werner T (1995) MatInd and MatInspector: new fast and versatile tools for detection of consensus matches in nucleotide sequence data. Nucleic Acids Res 23:4878–4884PubMedCrossRef
35.
Hannenhalli S, Kaestner KH (2009) The evolution of Fox genes and their role in development and disease. Nat Rev Genet 10:233–240PubMedCrossRef
36.
Friedman JR, Kaestner KH (2006) The Foxa family of transcription factors in development and metabolism. Cell Mol Life Sci 63:2317–2328PubMedCrossRef
37.
Overdier DG, Porcella A, Costa RH (1994) The DNA-binding specificity of the hepatocyte nuclear factor 3/forkhead domain is influenced by amino-acid residues adjacent to the recognition helix. Mol Cell Biol 14:2755–2766PubMedCrossRef
38.
Tuteja G, White P, Schug J, Kaestner KH (2009) Extracting transcription factor targets from ChIP-Seq data. Nucleic Acids Res 37:e113PubMedCrossRef
39.
Sparso T, Andersen G, Nielsen T et al (2008) The GCKR rs780094 polymorphism is associated with elevated fasting serum triacylglycerol, reduced fasting and OGTT-related insulinaemia, and reduced risk of type 2 diabetes. Diabetologia 51:70–75PubMedCrossRef
40.
Orho-Melander M, Melander O, Guiducci C et al (2008) Common missense variant in the glucokinase regulatory protein gene is associated with increased plasma triglyceride and C-reactive protein but lower fasting glucose concentrations. Diabetes 57:3112–3121PubMedCrossRef
41.
Pare G, Chasman DI, Parker AN et al (2008) Novel association of HK1 with glycated hemoglobin in a non-diabetic population: a genome-wide evaluation of 14,618 participants in the Women’s Genome Health Study. PLoS Genetics 4:e1000312PubMedCrossRef
42.
Soranzo N, Sanna S, Wheeler E et al (2010) Common variants at ten genomic loci influence hemoglobin A1C levels via glycemic and non-glycemic pathways. Diabetes 59:3229–3239PubMedCrossRef
43.
Lyssenko V, Lupi R, Marchetti P et al (2007) Mechanisms by which common variants in the TCF7L2 gene increase risk of type 2 diabetes. J Clin Invest 117:2155–2163PubMedCrossRef
44.
Gaulton KJ, Nammo T, Pasquali L et al (2010) A map of open chromatin in human pancreatic islets. Nat Genet 42:255–259PubMedCrossRef
45.
46.
Solis AS, Shariat N, Patton JG (2008) Splicing fidelity, enhancers, and disease. Front Biosci 13:1926–1942PubMedCrossRef
47.
Dos Santos C, Bougneres P, Fradin D (2009) An SNP in a methylatable Foxa2 binding site of the G6PC2 promoter is associated with insulin secretion in vivo and increased promoter activity in vitro. Diabetes 58:489–492PubMedCrossRef
48.
Gao N, Le Lay J, Qin W et al (2010) Foxa1 and Foxa2 maintain the metabolic and secretory features of the mature beta-cell. Mol Endocrinol 24:1594–1604PubMedCrossRef
49.
Taneera J, Lang S, Sharma A et al (2012) A systems genetics approach identifies genes and pathways for type 2 diabetes in human islets. Cell Metab 16:122–134PubMedCrossRef
50.
O’Brien RM, Noisin EL, Suwanichkul A et al (1995) Hepatic nuclear factor 3- and hormone-regulated expression of the phosphoenolpyruvate carboxykinase and insulin-like growth factor-binding protein 1 genes. Mol Cell Biol 15:1747–1758PubMed
Metadata
Title
Multiple functional polymorphisms in the G6PC2 gene contribute to the association with higher fasting plasma glucose levels
Authors
D. A. Baerenwald
A. Bonnefond
N. Bouatia-Naji
B. P. Flemming
O. C. Umunakwe
J. K. Oeser
L. D. Pound
N. L. Conley
S. Cauchi
S. Lobbens
E. Eury
B. Balkau
O. Lantieri
P. K. Dadi
D. A. Jacobson
P. Froguel
R. M. O’Brien
MAGIC Investigators
Publication date
01-06-2013
Publisher
Springer-Verlag
Published in
Diabetologia / Issue 6/2013
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-013-2875-3

Other articles of this Issue 6/2013

Diabetologia 6/2013 Go to the issue

Article

Adipose tissue hypoxia induces inflammatory M1 polarity of macrophages in an HIF-1α-dependent and HIF-1α-independent manner in obese mice

Commentary

Should more attention be paid to school performance in children and adolescents with diabetes?

Article

Oleate prevents saturated-fatty-acid-induced ER stress, inflammation and insulin resistance in skeletal muscle cells through an AMPK-dependent mechanism

Article

Otubain 2 is a novel promoter of beta cell survival as revealed by siRNA high-throughput screens of human pancreatic islets

Article

Systematic evaluation of validated type 2 diabetes and glycaemic trait loci for association with insulin clearance

Article

Distinct and opposing roles for the phosphatidylinositol 3-OH kinase catalytic subunits p110α and p110β in the regulation of insulin secretion from rodent and human beta cells
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits