The gene responsible for Dyggve-Melchior-Clausen syndrome encodes a novel peripheral membrane protein dynamically associated with the Golgi apparatus.

Dyggve-Melchior-Clausen dysplasia (DMC) is a rare inherited dwarfism with severe mental retardation due to mutations in the DYM gene which encodes Dymeclin, a 669-amino acid protein of yet unknown function. Despite a high conservation across species and several predicted transmembrane domains, Dymeclin could not be ascribed to any family of proteins. Here we show, using in situ hybridization, that DYM is widely expressed in human embryos, especially in the cortex, the hippocampus and the cerebellum. Both the endogenous and the recombinant protein fused to green fluorescent protein co-localized with Golgi apparatus markers. Electron microscopy revealed that Dymeclin associates with the Golgi apparatus and with transitional vesicles of the reticulum-Golgi interface. Moreover, permeabilization assays revealed that Dymeclin is not a transmembrane but a peripheral protein of the Golgi apparatus as it can be completely released from the Golgi after permeabilization of the plasma membrane. Time lapse confocal microscopy experiments on living cells further showed that the protein shuttles between the cytosol and the Golgi apparatus in a highly dynamic manner and recognizes specifically a subset of mature Golgi membranes. Finally, we found that DYM mutations associated with DMC result in mis-localization and subsequent degradation of Dymeclin. These data indicate that DMC results from a loss-of-function of Dymeclin, a novel peripheral membrane protein which shuttles rapidly between the cytosol and mature Golgi membranes and point out a role of Dymeclin in cellular trafficking.

[1]  H. Ruley,et al.  Dyggve–Melchior–Clausen syndrome: Chondrodysplasia resulting from defects in intracellular vesicle traffic , 2008, Proceedings of the National Academy of Sciences.

[2]  A. Linstedt,et al.  GRASP55 regulates Golgi ribbon formation. , 2008, Molecular biology of the cell.

[3]  Patrick Bouthemy,et al.  Non-parametric regression for patch-based fluorescence microscopy image sequence denoising , 2008, 2008 5th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[4]  C. Rabouille,et al.  dGRASP-mediated noncanonical integrin secretion is required for Drosophila epithelial remodeling. , 2008, Developmental cell.

[5]  J. Christopher Fromme,et al.  The genetic basis of a craniofacial disease provides insight into COPII coat assembly. , 2007, Developmental cell.

[6]  Patrick Bouthemy,et al.  Space-Time Adaptation for Patch-Based Image Sequence Restoration , 2007, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[7]  J. Christopher Fromme,et al.  Cranio-lenticulo-sutural dysplasia is caused by a SEC23A mutation leading to abnormal endoplasmic-reticulum-to-Golgi trafficking , 2006, Nature Genetics.

[8]  F. Lanni,et al.  GM130 and GRASP65-dependent lateral cisternal fusion allows uniform Golgi-enzyme distribution , 2006, Nature Cell Biology.

[9]  R. Trembath,et al.  Genomic duplication in Dyggve Melchior Clausen syndrome, a novel mutation mechanism in an autosomal recessive disorder , 2005, Journal of Medical Genetics.

[10]  K. Howell,et al.  GMx33 associates with the trans-Golgi matrix in a dynamic manner and sorts within tubules exiting the Golgi. , 2005, Molecular biology of the cell.

[11]  V. Malhotra,et al.  The Golgi-associated protein GRASP65 regulates spindle dynamics and is essential for cell division. , 2005, Molecular biology of the cell.

[12]  Ken Sato COPII coat assembly and selective export from the endoplasmic reticulum. , 2004, Journal of biochemistry.

[13]  Robert W. Taylor,et al.  NDUFS6 mutations are a novel cause of lethal neonatal mitochondrial complex I deficiency. , 2004, The Journal of clinical investigation.

[14]  A. Munnich,et al.  Recent advances in Dyggve-Melchior-Clausen syndrome. , 2004, Molecular genetics and metabolism.

[15]  M. Shaw,et al.  Human wild-type SEDL protein functionally complements yeast Trs20p but some naturally occurring SEDL mutants do not. , 2003, Gene.

[16]  B. Goud,et al.  Recombinant Antibodies Against Subcellular Fractions Used to Track Endogenous Golgi Protein Dynamics in Vivo , 2003, Traffic.

[17]  J. Gécz,et al.  Spondyloepiphyseal dysplasia tarda (SEDL, MIM #313400) , 2003, European Journal of Human Genetics.

[18]  M. Koenig,et al.  Frataxin deficiency in pancreatic islets causes diabetes due to loss of beta cell mass. , 2003, The Journal of clinical investigation.

[19]  A. Munnich,et al.  Mutations in a novel gene Dymeclin (FLJ20071) are responsible for Dyggve-Melchior-Clausen syndrome. , 2003, Human molecular genetics.

[20]  D. Rimoin,et al.  Mental retardation and abnormal skeletal development (Dyggve-Melchior-Clausen dysplasia) due to mutations in a novel, evolutionarily conserved gene. , 2003, American journal of human genetics.

[21]  Andrea C. Pfeifer,et al.  Dissection of COPI and Arf1 dynamics in vivo and role in Golgi membrane transport , 2002, Nature.

[22]  T. Casoli,et al.  Mapping of Mitochondrial Metabolic Competence by Cytochrome Oxidase and Succinic Dehydrogenase Cytochemistry , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[23]  P. Arvan,et al.  Disease models provide insights into post-golgi protein trafficking, localization and processing. , 1999, Current opinion in cell biology.

[24]  J. Gécz,et al.  Identification of the gene (SEDL) causing X-linked spondyloepiphyseal dysplasia tarda , 1999, Nature Genetics.

[25]  William E. Balch,et al.  Integration of endoplasmic reticulum signaling in health and disease , 1999, Nature Medicine.

[26]  Simon C Watkins,et al.  The distribution and translocation of the G protein ADP-ribosylation factor 1 in live cells is determined by its GTPase activity. , 1998, Journal of cell science.

[27]  P. Arvan,et al.  Endocrinopathies in the family of endoplasmic reticulum (ER) storage diseases: disorders of protein trafficking and the role of ER molecular chaperones. , 1998, Endocrine reviews.

[28]  G. Raposo,et al.  Accumulation of Major Histocompatibility Complex Class Ii Molecules in Mast Cell Secretory Granules and Their Release upon Degranulation Generation of Bmmcs Preparation of B Cells Reagents and Monoclonal Antibodies (mabs) Immunofluorescence Staining and Confocal Microscopy Pulse-chase# Labeling and , 2022 .

[29]  T. Kurokawa,et al.  Dyggve-Melchior-Clausen syndrome without mental retardation (Smith-McCort dysplasia): morphological findings in the growth plate of the iliac crest. , 1997, American journal of medical genetics.

[30]  L. Hedstrom,et al.  Isolation and Characterization of Mycophenolic Acid-resistant Mutants of Inosine-5′-monophosphate Dehydrogenase* , 1997, The Journal of Biological Chemistry.

[31]  P. Beighton,et al.  Dyggve-Melchior-Clausen syndrome. , 1990, Journal of medical genetics.

[32]  H. Kresse,et al.  Dyggve-Melchior-Clausen syndrome: normal degradation of proteodermatan sulfate, proteokeratan sulfate and heparan sulfate. , 1984, Clinica chimica acta; international journal of clinical chemistry.

[33]  J. Lippincott-Schwartz,et al.  The fluorescence protease protection (FPP) assay to determine protein localization and membrane topology , 2006, Nature Protocols.

[34]  D. Wilkinson In situ hybridization: a practical approach , 1998 .

[35]  C. Dietrich,et al.  Dyggve-Melchior-Clausen syndrome: genetic studies and report of affected sibs. , 1979, American journal of medical genetics.