Genomics Inform.  2014 Dec;12(4):240-246. 10.5808/GI.2014.12.4.240.

Identification and Functional Characterization of P159L Mutation in HNF1B in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5)

• Kim EK
• Lee JS
• Cheong HI
• Chung SS
• Kwak SH
• Park KS

• KMID: 2391503
• DOI: http://doi.org/10.5808/GI.2014.12.4.240

Abstract

Mutation in HNF1B, the hepatocyte nuclear factor-1beta (HNF-1beta) gene, results in maturity-onset diabetes of the young (MODY) 5, which is characterized by gradual impairment of insulin secretion. However, the functional role of HNF-1beta in insulin secretion and glucose metabolism is not fully understood. We identified a family with early-onset diabetes that fulfilled the criteria of MODY. Sanger sequencing revealed that a heterozygous P159L (CCT to CTT in codon 159 in the DNA-binding domain) mutation in HNF1B was segregated according to the affected status. To investigate the functional consequences of this HNF1B mutation, we generated a P159L HNF1B construct. The wild-type and mutant HNF1B constructs were transfected into COS-7 cells in the presence of the promoter sequence of human glucose transporter type 2 (GLUT2). The luciferase reporter assay revealed that P159L HNF1B had decreased transcriptional activity compared to wild-type (p < 0.05). Electrophoretic mobility shift assay showed reduced DNA binding activity of P159L HNF1B. In the MIN6 pancreatic beta-cell line, overexpression of the P159L mutant was significantly associated with decreased mRNA levels of GLUT2 compared to wild-type (p < 0.05). However, INS expression was not different between the wild-type and mutant HNF1B constructs. These findings suggests that the impaired insulin secretion in this family with the P159L HNF1B mutation may be related to altered GLUT2 expression in beta-cells rather than decreased insulin gene expression. In conclusion, we have identified a Korean family with an HNF1B mutation and characterized its effect on the pathogenesis of diabetes.

Keyword

glucose transporter type 2; hepatocyte nuclear factor-1beta; point mutation; type 2 diabetes mellitus

MeSH Terms

Animals
Codon
COS Cells
Diabetes Mellitus, Type 2*
DNA
Electrophoretic Mobility Shift Assay
Gene Expression
Glucose
Glucose Transporter Type 2
Hepatocyte Nuclear Factor 1-beta
Humans
Insulin
Luciferases
Metabolism
Point Mutation
RNA, Messenger
Codon
DNA
Glucose
Glucose Transporter Type 2
Hepatocyte Nuclear Factor 1-beta
Insulin
Luciferases
RNA, Messenger