Kniest dysplasia is a heritable chondrodysplasia that severely affects skeletal growth. Recent evidence suggests that the etiology is based on mutations in COL2A1, the gene for
collagen type II. We report the detection and partial characterization of an identical defect in
type II collagen in two unrelated patients with
Kniest dysplasia. Analysis of
cyanogen bromide (CB)-digested cartilage samples from both probands by SDS-PAGE revealed an abnormal band for
peptide alpha 1(II)CB12. The
peptide was purified and digested with
endoproteinase Asp-N. Fragments unique to the Kniest tissues were identified by reverse-phase high-pressure liquid chromatography and by sequence analysis. The results established a deletion of
amino acids 102-108 of the alpha 1(II) triple-helical domain, which disrupted the (gly-X-Y)n repeat needed for helix formation. This was confirmed by sequence analysis of
DNA amplified from both probands, revealing the molecular basis to be a single
nucleotide mutation at a CpG dinucleotide (GCG-->GTG) in the
codon for
alanine 102. The mutation created a new
splice donor site, which would account for the absence of the last seven
amino acids from the 3' end of exon 12 in alpha 1(II)CB12. Light and electron micrographs of the probands' cartilage showed the perilacunar foamy matrix ("Swiss cheese") characteristic of
Kniest dysplasia and chondrocytes containing dilated rough endoplasmic reticulum, which earlier studies had shown were filled with
type II procollagen. These two cases strengthen the concept that
Kniest dysplasia is based on mutations of COL2A1 and belongs within the broad spectrum of chondrodysplasias caused by type II collagenopathies.