6 MUTATIONS IN THE GLUCOKINASE GENE IDENTIFIED IN MODY BY USING A NONRADIOACTIVE SENSITIVE SCREENING TECHNIQUE

We have reported that 56% of French families with maturity-onset diabe tes of the young (MODY) carry a mutation in the glucokinase gene (GCK) . Therefore, we have established a quick and sensitive nonradioactive technique (with the PhastSystem(TM) based on single-strand conformatio n polymorphism [SSCP] analysis) to routinely screen the 12 exons of GC K for mutations. We have studied GCK in 12 young hyperglycemic patient s with a strong family history of type II diabetes. SSCP variants were observed in 6 of those 12 patients (50%), which cosegregated with dia betes in five families where DNA hom additional members was available. Direct sequencing identified a 10-bp (base pair) deletion in exon 3; a 33-bp deletion at the exon 5/intron 5 junction, including the two co nsensus bases (GT) of the donor splice site; a nonsense mutation in ex on 5 (Arg(186) -> Stop) in a Black-African family, which has been iden tified previously in a Caucasian family; and three missense mutations: Thr(209) -> Met(209) in exon 6, Gly(261) -> Glu(261) in exon 7, and A rg(36) -> Trp(36) in exon 2. The missense mutation in exon 2 was found only in the second and third generation of the tested family but not in the first. To our knowledge, this is the first time that a de novo mutation of GCK is reported within a family. All six families carrying a mutation in GCK were typical MODY and most of their affected member s had a mild form of diabetes. This nonradioactive SSCP technique may be useful to routinely diagnose glucokinase deficiency, which is an im portant cause of hyperglycemia among young type II diabetic patients.