Human Molecular Genetics Advance Access originally published online on July 29, 2009
Human Molecular Genetics 2009 18(21):4013-4021; doi:10.1093/hmg/ddp345
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A gradient of ROR2 protein stability and membrane localization confers brachydactyly type B or Robinow syndrome phenotypes
1 Max Planck-Institute for Molecular Genetics, FG Development & Disease, Berlin, Germany, 2 Institute for Medical Genetics, Charité and 3 Berlin-Brandenburg Center for Regenerative Therapies (BCRT), University Medicine Berlin, Berlin, Germany and 4 Genetic Unit, DGHA, Ministry of Health, Muscat, Sultanate of Oman
* To whom correspondence should be addressed at: Institute for Medical Genetics, Charité, University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin, Germany. Tel: +49 30450569121; Fax: +49 30450569915; Email: stefan.mundlos{at}charite.de
Received April 22, 2009; Revised July 7, 2009; Accepted July 22, 2009
Mutations in ROR2 cause dominant brachydactyly type B (BDB1) or recessive Robinow syndrome (RRS), each characterized by a distinct combination of phenotypic features. We here report a novel nonsense mutation in ROR2 (c.1324C>T; p.R441X) causing intracellular protein truncation in a patient exhibiting features of RRS in conjunction with severe recessive brachydactyly. The mutation is located at the same position as a previously described frame shift mutation causing dominant BDB1. To investigate the apparent discrepancy in phenotypic outcome, we analysed ROR2 protein stability and distribution in stably transfected cell lines expressing exact copies of several human RRS and BDB1 intracellular mutations. RRS mutant proteins were less abundant and retained intracellularly, although BDB1 mutants were stable and predominantly located at the cell membrane. The p.R441X mutation showed an intermediate pattern with membrane localization but also high endoplasmic reticulum retention. Furthermore, we observed a correlation between the severity of BDB1, the location of the mutation, and the amount of membrane-associated ROR2. Membrane protein fraction quantification revealed a gradient of distribution and stability correlating with the clinical phenotypes. This gradual model was confirmed by crossing mouse models for RRS and BDB1, yielding double heterozygous animals that exhibited an intermediate phenotype. We propose a model in which the RRS versus the BDB1 phenotype is determined by the relative degree of protein retention/degradation and the amount of mutant protein reaching the plasma membrane.