title: Characterization of a naturally occurring mutation (L107I) in the HNF1 alpha (MODY3) gene.
abstract: OBJECTIVE
  Maturity onset diabetes of the young type 3 (MODY3) is a monogenic form of diabetes mellitus caused by mutations in the gene encoding for hepatocyte nuclear factor 1 alpha, HNF1 alpha. In this study we have examined the in vivo and in vitro effects of a mutation (L107I) outside the DNA binding and dimerization domains in the N terminal part of the HNF1 alpha gene.
  
  
  METHODS
  Beta-cell function of the affected family members was assessed by an oral glucose tolerance test. Functional tests were carried out to explain the role of the mutation in vitro by transcriptional activity assay, Western blotting, DNA-binding assays and subcellular localization experiments.
  
  
  RESULTS
  Affected family members showed an 86% decreased insulin response to glucose when compared to age-matched healthy control subjects. In vitro the mutation showed a 79% decrease in transcriptional activity as compared to wild type HNF1 alpha in HeLa cells lacking HNF1 alpha. The transcriptional activity was not suppressed when the mutant was co-expressed with wild type HNF1 alpha suggesting that the decreased activity was not mediated by a dominant negative mechanism. The L107I/HNF1alpha protein showed normal nuclear targeting but impaired binding to an HNF1 alpha consensus sequence.
  
  
  CONCLUSIONS
  Our results suggest that the L107I substitution represents a MODY3 mutation which impairs beta-cell function by a loss-of-function mechanism.
authors.affiliation: Department of Endocrinology, Wallenberg Laboratory, Malmö University Hospital, Sweden.
authors.forename: C
authors.initials: C
authors.lastname: Cervin
authors.forename: M
authors.initials: M
authors.lastname: Orho-Melander
authors.forename: M
authors.initials: M
authors.lastname: Ridderstråle
authors.forename: M
authors.initials: M
authors.lastname: Lehto
authors.forename: S
authors.initials: S
authors.lastname: Barg
authors.forename: L
authors.initials: L
authors.lastname: Groop
authors.forename: C M
authors.initials: CM
authors.lastname: Cilio
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution; D001692:Biological Transport / Q000235:genetics; D001786:Blood Glucose / Q000032:analysis; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D019281:Dimerization; D005260:Female; D051537:Hepatocyte Nuclear Factor 1; D051538:Hepatocyte Nuclear Factor 1-alpha; D051539:Hepatocyte Nuclear Factor 1-beta; D006579:Heterozygote; D006801:Humans; D007328:Insulin / Q000097:blood; D007532:Isoleucine; D007930:Leucine; D008297:Male; D009154:Mutation / Q000235:genetics*; D009687:Nuclear Proteins; D010375:Pedigree; D017434:Protein Structure, Tertiary / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D001786:Blood Glucose; D004268:DNA-Binding Proteins; C496487:HNF1A protein, human; C496495:HNF1B protein, human; D051538:Hepatocyte Nuclear Factor 1-alpha; D007328:Insulin; D009687:Nuclear Proteins; D014157:Transcription Factors; D007532:Isoleucine; D051537:Hepatocyte Nuclear Factor 1; D051539:Hepatocyte Nuclear Factor 1-beta; D004247:DNA; D007930:Leucine