MMP13 mutation causes spondyloepimetaphyseal dysplasia, Missouri type (SEMDMO)
Ann M. Kennedy, … , Michael P. Whyte, Rajesh V. Thakker
Ann M. Kennedy, … , Michael P. Whyte, Rajesh V. Thakker
Published October 3, 2005
Citation Information: J Clin Invest. 2005;115(10):2832-2842. https://doi.org/10.1172/JCI22900.
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Research Article Bone biology

MMP13 mutation causes spondyloepimetaphyseal dysplasia, Missouri type (SEMDMO)

Abstract

MMPs, which degrade components of the ECM, have roles in embryonic development, tissue repair, cancer, arthritis, and cardiovascular disease. We show that a missense mutation of MMP13 causes the Missouri type of human spondyloepimetaphyseal dysplasia (SEMDMO), an autosomal dominant disorder characterized by defective growth and modeling of vertebrae and long bones. Genome-wide linkage analysis mapped SEMDMO to a 17-cM region on chromosome 11q14.3–23.2 that contains a cluster of 9 MMP genes. Among these, MMP13 represented the best candidate for SEMDMO, since it preferentially degrades collagen type II, abnormalities of which cause skeletal dysplasias that include Strudwick type SEMD. DNA sequence analysis revealed a missense mutation, F56S, that substituted an evolutionarily conserved phenylalanine residue for a serine in the proregion domain of MMP13. We predicted, by modeling MMP13 structure, that this F56S mutation would result in a hydrophobic cavity with misfolding, autoactivation, and degradation of mutant protein intracellularly. Expression of wild-type and mutant MMP13s in human embryonic kidney cells confirmed abnormal intracellular autoactivation and autodegradation of F56S MMP13 such that only enzymatically inactive, small fragments were secreted. Thus, the F56S mutation results in deficiency of MMP13, which leads to the human skeletal developmental anomaly of SEMDMO.

Authors

Ann M. Kennedy, Masaki Inada, Stephen M. Krane, Paul T. Christie, Brian Harding, Carlos López-Otín, Luis M. Sánchez, Anna A.J. Pannett, Andrew Dearlove, Claire Hartley, Michael H. Byrne, Anita A.C. Reed, M. Andrew Nesbit, Michael P. Whyte, Rajesh V. Thakker

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Figure 5

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Detection of mutation in exon 2 by restriction enzyme analysis. (A) DNA ...
Detection of mutation in exon 2 by restriction enzyme analysis. (A) DNA sequence analysis of the affected female III.7 revealed a T→C transition at codon 56. The wild-type sequence at codon 56 is TTC encoding a phenylalanine (Phe, F) residue, whereas the mutant (Mu) is TCC encoding for a serine (Ser, S) residue. This missense mutation resulted in the gain of an MspI restriction site (C/CGGC), which facilitated its detection in the family with SEMDMO (B). (C) The restriction enzyme map of the PCR products shows that the wild-type DNA sequence results in a 269-bp product, whereas the mutant, which has one MspI site, is associated with cleaved PCR products of 118 bp and 151 bp. Analysis of the SEMDMO family (Figure 2) revealed that the affected members (II.4, III.7, III.9, IV.6, and IV.7; shown [filled symbols]) were heterozygous for the F56S mutation and that the unaffected members (II.5, III.5, III.6, III.10, and III.11; shown [open symbols]) were homozygous for the wild-type sequence. Standard size markers (S), in the form of a 100-bp ladder, are shown. Cosegregation of this mutation with the disease in the family (Figure 2), in conjunction with its absence in 110 alleles from 55 normal individuals (N1 and N2 are shown) indicates that it is not a common DNA sequence polymorphism.