RESUMEN
Winchester syndrome (WS, MIM #277950) is an extremely rare autosomal recessive skeletal dysplasia characterized by progressive joint destruction and osteolysis. To date, only one missense mutation in MMP14, encoding the membrane-bound matrix metalloprotease 14, has been reported in WS patients. Here, we report a novel hypomorphic MMP14 p.Arg111His (R111H) allele, associated with a mitigated form of WS. Functional analysis demonstrated that this mutation, in contrast to previously reported human and murine MMP14 mutations, does not affect MMP14's transport to the cell membrane. Instead, it partially impairs MMP14's proteolytic activity. This residual activity likely accounts for the mitigated phenotype observed in our patients. Based on our observations as well as previously published data, we hypothesize that MMP14's catalytic activity is the prime determinant of disease severity. Given the limitations of our in vitro assays in addressing the consequences of MMP14 dysfunction, we generated a novel mmp14a/b knockout zebrafish model. The fish accurately reflected key aspects of the WS phenotype including craniofacial malformations, kyphosis, short-stature and reduced bone density owing to defective collagen remodeling. Notably, the zebrafish model will be a valuable tool for developing novel therapeutic approaches to a devastating bone disorder.
Asunto(s)
Anomalías Múltiples/genética , Contractura/genética , Opacidad de la Córnea/genética , Anomalías Craneofaciales/genética , Trastornos del Crecimiento/genética , Metaloproteinasa 14 de la Matriz/genética , Osteólisis/genética , Osteoporosis/genética , Anomalías Múltiples/fisiopatología , Alelos , Animales , Dominio Catalítico/genética , Contractura/fisiopatología , Opacidad de la Córnea/fisiopatología , Anomalías Craneofaciales/fisiopatología , Técnicas de Inactivación de Genes , Trastornos del Crecimiento/fisiopatología , Humanos , Ratones , Osteólisis/fisiopatología , Osteoporosis/fisiopatología , Fenotipo , Pez CebraRESUMEN
Correct positioning of the cell division machinery is key to genome stability. Schizosaccharomyces pombe is an attractive organism to study cytokinesis as it, like higher eukaryotes, divides using a contractile actomyosin ring. In S. pombe, many actomyosin ring components assemble at the medial cortex into node-like structures before coalescing into a ring [1, 2]. Assembly of cytokinetic nodes requires Mid1p, which recruits IQGAP-related Rng2p to the division site, after which other node components accumulate at the division site in a characteristic sequence [3-6]. How cytokinetic nodes assemble, whether the order of assembly of ring components is important, and whether Mid1p solely participates in ring positioning are poorly understood. Here, we show that synthetic targeting of IQGAP-related Rng2p, formin-Cdc12p, and myosin II (Myo2p) restores medial division in mid1 mutants, suggesting that ring proteins need not assemble at the division site in an invariant order. Unlike in wild-type cells, actomyosin rings in cells rewired to divide medially in the absence of Mid1p assemble late in anaphase. Furthermore, the rewiring process affects the ability of the actomyosin ring to track the nucleus upon perturbation of nuclear position. Our work reveals the power of synthetic rewiring studies in deciphering roles performed by multifunctional proteins.
Asunto(s)
Citocinesis , Proteínas de Schizosaccharomyces pombe/genética , Schizosaccharomyces/crecimiento & desarrollo , Schizosaccharomyces/genética , Reacción en Cadena de la Polimerasa , Schizosaccharomyces/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismoRESUMEN
In many eukaryotes, cytokinesis requires an actomyosin contractile ring that is crucial for cell constriction and new membrane organization. Two studies in this issue (Onishi et al. 2013. J. Cell Biol. http://dx.doi.org.10.1083/jcb.201302001 and Atkins et al. 2013. J. Cell Biol. http://dx.doi.org.10.1083/jcb.201301090) establish that precise activation and/or inactivation of Rho1 and Cdc42 GTPases is important for the correct order and successful completion of events downstream of actomyosin ring constriction in budding yeast.