Identification and functional characterization of a novel mutation of hepatocyte nuclear factor-1α gene in a Korean family with MODY3

K. A. Kim, K. Kang, Y. I. Chi, I. Chang, M. K. Lee, K. W. Kim, S. E. Shoelson, M. S. Lee

Research output: Contribution to journalArticlepeer-review

19 Scopus citations


Aims/hypothesis. After screening 16 Korean families with early onset Type 2 diabetes in search for hepatocyte nuclear factor (HNF)-1α gene mutation, we identified a novel missense mutation (R263L) associated with MODY phenotype. We studied the biological characteristics of the mutation and the potential functional consequences based on the crystallographic structure of HNF-1α in complex with DNA. Methods. DNA from subjects with a familial form of early onset diabetes was isolated and HNF-1α was sequenced. The R263L substitution was generated by PCR-based sited-directed mutagenesis. Functional and biochemical studies were conducted by reporter assay using glucose-transporter type 2 (GLUT2) or insulin promoters and electrophoretic mobility shift assay, respectively. Results. Transfection of wild-type HNF-1α increased the reporter activities of GLUT2 and insulin promoters in NIH3T3 and SK-Hep1 cells, while R263L mutant was defective in transactivation of those promoters. Both wild-type HNF-1α and R263L mutant could not transactivate GLUT2 and insulin promoters in MIN6N8 insulinoma cells. R263L mutant had a defective cooperation with its heterodimeric partner HNF-1β or coactivator p300. R263L mutant protein displayed greatly reduced DNA binding ability, despite its comparable protein stability to the wild-type HNF-1α. Conclusion/interpretation. These results suggest that the mutation of HNF-1α at codon 263 from arginine to leucine leads to the development of MODY3 through decreased insulin production and defective glucose sensing. These findings are in good agreement with the crystal structure in which R263 makes hydrogen bonds with phosphorus atoms of DNA backbone to mediate the stable binding of HNF-1α homeodomain to the promoter.

Original languageEnglish (US)
Pages (from-to)721-727
Number of pages7
Issue number5
StatePublished - May 1 2003

Bibliographical note

Funding Information:
Acknowledgements. This work was supported by Science Research Centre Grants from the Korea Science & Engineering Foundation and Health Planning Technology & Evaluation Board Grants (02-PJ1-PG1-CH04-0001) (M.-S.L.). This work was also supported by NIH grant R01 DK43123 (S.E.S.) and a fellowship from the Mary K. Iacocca Foundation (Y.-I.C.). M.-S. Lee is an awardee of the National Research Laboratory Grants from the Korea Institute of Science & Technology Evaluation and Planning (2000-N-NL-01-C-232). We thank Dr. J. Takeda (Gunma University) for generously providing human HNF-1α expression vector (pCMV6b-hHNF-1α), HNF-1β expression vector and pGL3-GT2. We are also grateful to Dr. K. Yamagata (Osaka University) for providing pGL3-Insulin construct and Dr. J.W. Lee (Pohang University of Science and Technology, Pohang, Korea) for providing p300. We are indebted to Dr. M.S. Nam for helpful discussion and Dr. B.-C. Oh for the production of R263L mutant construct.


  • Glucose transporter type 2
  • HNF-1α
  • HNF-1β
  • Insulin
  • MODY
  • Mutation
  • p300


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