Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5)
Mutation in HNF1B, the hepatocyte nuclear factor-1β (HNF-1β) 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-1β in insulin secretion and glucose metabolism is not fully understood....
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BioMed Central
2014-12-01
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| Series: | Genomics & Informatics |
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| Online Access: | http://genominfo.org/upload/pdf/gni-12-240.pdf |
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| author | Eun Ky Kim Ji Seon Lee Hae Il Cheong Sung Soo Chung Soo Heon Kwak Kyong Soo Park |
| author_facet | Eun Ky Kim Ji Seon Lee Hae Il Cheong Sung Soo Chung Soo Heon Kwak Kyong Soo Park |
| author_sort | Eun Ky Kim |
| collection | DOAJ |
| description | Mutation in HNF1B, the hepatocyte nuclear factor-1β (HNF-1β) 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-1β 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 β-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 β-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. |
| format | Article |
| id | doaj-art-17838f89745b4dac9d4683a373940255 |
| institution | Kabale University |
| issn | 1598-866X 2234-0742 |
| language | English |
| publishDate | 2014-12-01 |
| publisher | BioMed Central |
| record_format | Article |
| series | Genomics & Informatics |
| spelling | doaj-art-17838f89745b4dac9d4683a3739402552025-02-02T02:22:59ZengBioMed CentralGenomics & Informatics1598-866X2234-07422014-12-0112424024610.5808/GI.2014.12.4.240113Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5)Eun Ky Kim0Ji Seon Lee1Hae Il Cheong2Sung Soo Chung3Soo Heon Kwak4Kyong Soo Park5Department of Internal Medicine, Seoul National University College of Medicine, Seoul 110-744, Korea.Department of Internal Medicine, Seoul National University College of Medicine, Seoul 110-744, Korea.Department of Pediatrics, Seoul National University Children's Hospital, Seoul 110-744, Korea.Department of Internal Medicine, Seoul National University College of Medicine, Seoul 110-744, Korea.Department of Internal Medicine, Seoul National University Hospital, Seoul 110-744, Korea.Department of Internal Medicine, Seoul National University College of Medicine, Seoul 110-744, Korea.Mutation in HNF1B, the hepatocyte nuclear factor-1β (HNF-1β) 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-1β 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 β-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 β-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.http://genominfo.org/upload/pdf/gni-12-240.pdfglucose transporter type 2hepatocyte nuclear factor-1βpoint mutationtype 2 diabetes mellitus |
| spellingShingle | Eun Ky Kim Ji Seon Lee Hae Il Cheong Sung Soo Chung Soo Heon Kwak Kyong Soo Park Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) Genomics & Informatics glucose transporter type 2 hepatocyte nuclear factor-1β point mutation type 2 diabetes mellitus |
| title | Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) |
| title_full | Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) |
| title_fullStr | Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) |
| title_full_unstemmed | Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) |
| title_short | Identification and Functional Characterization of P159L Mutation in in a Family with Maturity-Onset Diabetes of the Young 5 (MODY5) |
| title_sort | identification and functional characterization of p159l mutation in in a family with maturity onset diabetes of the young 5 mody5 |
| topic | glucose transporter type 2 hepatocyte nuclear factor-1β point mutation type 2 diabetes mellitus |
| url | http://genominfo.org/upload/pdf/gni-12-240.pdf |
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