DOCSLIB.ORG

Dystrobrevin and Desmin

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Dystrobrevin and Desmin Desmuslin, an intermediate filament protein that interacts with ␣-dystrobrevin and desmin Yuji Mizuno*, Terri G. Thompson*, Jeffrey R. Guyon*, Hart G. W. Lidov*, Melissa Brosius*, Michihiro Imamura†, Eijiro Ozawa†, Simon C. Watkins‡, and Louis M. Kunkel*§ *Howard Hughes Medical Institute͞Division of Genetics, Children’s Hospital and Harvard Medical School, Boston, MA 02115; †National Institute of Neuroscience, National Center for Neurology and Psychiatry, 4-1-1 Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan; and ‡Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, PA 15261 Contributed by Louis M. Kunkel, March 28, 2001 Dystrobrevin is a component of the dystrophin-associated protein (19), the rabbit 94-kDa protein (A0) (20), and ␤-dystrobrevin complex and has been shown to interact directly with dystrophin, (21). ␣-Dystrobrevin 1 has a unique C-terminal region with ␣1-syntrophin, and the sarcoglycan complex. The precise role of multiple sites for tyrosine phosphorylation and is highly ex- ␣-dystrobrevin in skeletal muscle has not yet been determined. To pressed in muscle and brain. This protein has two predicted study ␣-dystrobrevin’s function in skeletal muscle, we used the ␣-helical coiled-coil motifs and has been shown to interact yeast two-hybrid approach to look for interacting proteins. Three directly with ␣1-syntrophin (16, 17) and dystrophin (12). The overlapping clones were identified that encoded an intermediate ␣-dystrobrevin 2 splice form is slightly different in that it lacks filament protein we subsequently named desmuslin (DMN). Se- the unique C-terminal region and thus would not be phosphor- quence analysis revealed that DMN has a short N-terminal domain, ylated. ␣-Dystrobrevin 3 has an alternatively spliced 3Ј end that a conserved rod domain, and a long C-terminal domain, all common is more truncated than that of ␣-dystrobrevin 2. ␣-Dystrobrevin features of type 6 intermediate filament proteins. A positive 4 and 5 have a different 5Ј start site relative to variants 1–3. interaction between DMN and ␣-dystrobrevin was confirmed with To better understand the role of ␣-dystrobrevin in skeletal an in vitro coimmunoprecipitation assay. By Northern blot analysis, muscle, we looked for interacting proteins by using the yeast we find that DMN is expressed mainly in heart and skeletal muscle, two-hybrid technique. We isolated three overlapping clones and although there is some expression in brain. Western blotting confirmed their interaction with dystrobrevin with in vitro co- detected a 160-kDa protein in heart and skeletal muscle. Immuno- immunoprecipitation (CoIP). A full-length cDNA clone of the fluorescent microscopy localizes DMN in a stripe-like pattern in interacting protein was isolated. This protein, desmuslin (DMN), longitudinal sections and in a mosaic pattern in cross sections of colocalizes to the Z-lines with desmin. Our results suggest that skeletal muscle. Electron microscopic analysis shows DMN colocal- DMN forms a linkage between desmin and extracellular matrix ized with desmin at the Z-lines. Subsequent coimmunoprecipita- and therefore provides an important structural support in tion experiments confirmed an interaction with desmin. Our find- muscle. ings suggest that DMN may serve as a direct linkage between the extracellular matrix and the Z-discs (through plectin) and may play Materials and Methods an important role in maintaining muscle cell integrity. Yeast Two-Hybrid Library Screening. ␣-Dystrobrevin cDNA (nucle- otides 7–1617) was inserted downstream of the Gal-4 DNA- he severe muscle wasting disorder, Duchenne muscular binding domain in the pGBT9 bait vector. A yeast two-hybrid Tdystrophy, is caused by abnormalities in the dystrophin gene cDNA library derived from human skeletal muscle was screened (1). The dystrophin protein is expressed in heart and skeletal for interacting proteins with the use of the Matchmaker two- muscle, where it is part of the dystrophin-associated protein hybrid system as described by the distributor (CLONTECH). complex. Dystrophin’s N-terminal domain binds to actin, In brief, transformation mixtures were spread on synthetic whereas the WW domain and the total cysteine-rich domain bind dropout͞ϪHis͞ϪLeu͞ϪTrp͞ϩ3-amino-1,2,4-triazole plates to ␤-dystroglycan (2), a component of the dystroglycan subcom- and incubated at 30°C until colonies appeared (6 days). Colonies plex. This subcomplex links to laminin, a major component of the able to grow on minimal plates were screened for ␤-galactosidase basal membrane, thereby forming the linkage between an intra- activity with the use of a filter-lift assay as described by the cellular protein, actin, and the extracellular matrix. manufacturer. Yeast DNA isolated from colonies positive for A second subcomplex of the dystrophin-associated protein ␤-galactosidase activity was used to electroporate Escherichia complex includes four transmembrane proteins (␣-, ␤-, ␥-, and coli to recover the interacting cDNA. The sequence of the ␦-sarcoglycan) (3). Each has been shown to be involved in interacting cDNA was analyzed on an ABI 373 or 377 automated different forms of limb-girdle muscular dystrophy (LGMD 2D, sequencer with the use of fluorescent dye terminator chemistry 2E, 2C, and 2F) (4–8). ␣-Sarcoglycan is a type 1 transmembrane (Applied Biosystems). protein and is expressed in heart and skeletal muscle. ␤-, ␥-, and ␦-sarcoglycans are type 2 transmembrane proteins containing a Phage cDNA Library Screening. A ␭gt 11 human skeletal muscle cluster of cysteine residues in their extracellular domains. These library (CLONTECH) was screened with a 152-bp hybridization four proteins form the sarcoglycan complex, which is thought to probe homologous to DMN’s 5Ј region (Fig. 2A, nucleotides Ϫ23 be involved in some type of signaling pathway (9). A third subcomplex of the dystrophin-associated protein complex involves ␣-dystrobrevin (10–12) and the syntrophins Abbreviations: CoIP, coimmunoprecipitation; DMN, desmuslin; IF, intermediate filament. (␣1, ␤1, and ␤2) (13–15). These intracellular proteins directly Data deposition: The sequence reported in this paper has been deposited in the GenBank bind to dystrophin (16, 17). In addition, the N-terminal region of database (accession no. AF359284). ␣-dystrobrevin associates with the sarcoglycan complex (18). §To whom reprint requests should be addressed at: Howard Hughes Medical Institute, Division of Genetics, Enders 570, Children’s Hospital, 300 Longwood Avenue, Boston, MA There are at least five different forms of ␣-dystrobrevin gener- ␣ 02115. E-mail: [email protected]. ated by alternative splicing. The largest splice variant -dystro- The publication costs of this article were defrayed in part by page charge payment. This brevin 1 shows sequence homology to the cysteine-rich and article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. C-terminal domains of dystrophin, the Torpedo 87-kDa protein §1734 solely to indicate this fact. 6156–6161 ͉ PNAS ͉ May 22, 2001 ͉ vol. 98 ͉ no. 11 www.pnas.org͞cgi͞doi͞10.1073͞pnas.111153298 Downloaded by guest on September 26, 2021 of the subcloned cDNAs also was expressed from the pFHR2 vector along with the rest of the protein. Proteins expressed from these vectors were termed FLAG 4C, FLAG 5D-1, or FLAG 5D-2 (Fig. 1B). We also made several truncated versions of DMN to identify the subregion that interacted with ␣-dystrobrevin. Constructs were prepared by PCR amplification, using full-length DMN as the template. The DMN PCR product covered the sequence from Ϫ3 to 1011 and was subcloned into pFHR2 to express FLAG 1A-1B-2A-2B (amino acids Ϫ1 to 337). Constructs 1A-1B-2A (amino acids Ϫ1 to 225), 2A-2B (amino acids 161– 337), 1B-2A (amino acids 62–225), C-terminal-1 (amino acids 336–933), and C-terminal-2 (amino acids 932-1253) were cloned in a similar fashion. Clone 4C inserted into pFHR2 was used to express FLAG 1A-1B (amino acids Ϫ34 to 153). Desmin was cloned by PCR amplification from total cellular RNA isolated from human skeletal muscle (CLONTECH). The Fig. 1. CoIP analysis of DMN clones with dystrobrevin. (A) Alignment of PCR product was subcloned into pMGT1 and sequenced. KIAA0353 (22) and DMN clones 4C, 5D, 5D-1, and 5D-2. IF designates a region with an intermediate filament signature. (B) When simultaneously expressed, Antibodies. Synthetic peptides corresponding to DMN-1 amino FLAG 5D-1 coimmunoprecipitates dystrobrevin (compare lane 5 to lane 4). FLAG 5D-2 and FLAG 4C do not coimmunoprecipitate dystrobrevin (lanes 6 acids 232–250 (C-QEAEALRREALGLEQLRAR) and and 7). Our controls show that the FLAG antibody specifically precipitates DMN-2 amino acids 1038–1053 (C-SLSRQRSPAPGSPDEE) proteins with the FLAG epitope (lanes 1–3) and precipitates background levels (Fig. 2A) were generated and used to inject New Zealand White of dystrobrevin (lane 4). * indicates 35S-labeled dystrobrevin. ϩ͞Ϫ indicates rabbits (Research Genetics, Huntsville, AL). The amino- the presence or absence of a particular construct. terminal cysteine is not part of the DMN sequence, but was added for subsequent affinity purification with the use of a SulfoLink coupling gel (Pierce). Anti-FLAG mAb was pur- to 129). The probe was labeled with the use of a Random Primer chased from Sigma. DNA Labeling System (GIBCO͞BRL) in the presence of 5 ␮Ci of [␣-32P]dCTP. Filters were prehybridized in hybridization In Vitro Transcription͞Translation and CoIP. Proteins encoding the buffer [5ϫ SSC (0.75 M NaCl͞0.075 M sodium citrate)͞50 mM various constructs were labeled with [35S]methionine with a TNT sodium phosphate (pH 7.4)͞1ϫ Denhardt’s solution (0.02% Quick Coupled Transcription͞Translation System (Promega). Ficoll [type 400]͞0.02% polyvinylpyrrolidone͞0.02% BSA)] for Depending on the experiment, the proteins were expressed 5.5 h at 65°C. The denatured probe (8.9 ϫ 105 cpm͞ml) was either individually or simultaneously. Five microliters of protein added to the hybridization buffer, and the filters were hybridized lysate was added to 25 ␮l of CoIP buffer [150 mM NaCl͞50 mM for 18 h at 65°C. Filters were washed in 2ϫ SSC and 0.1% SDS Tris⅐HCl (pH 7.4)͞1% Nonidet P-40͞Protease Inhibitor Mixture for 30 min at 55°C with several changes of washing solution.
Load more

Recommended publications
  • Genetic Mutations and Mechanisms in Dilated Cardiomyopathy
    Genetic mutations and mechanisms in dilated cardiomyopathy Elizabeth M. McNally, … , Jessica R. Golbus, Megan J. Puckelwartz J Clin Invest. 2013;123(1):19-26. https://doi.org/10.1172/JCI62862. Review Series Genetic mutations account for a significant percentage of cardiomyopathies, which are a leading cause of congestive heart failure. In hypertrophic cardiomyopathy (HCM), cardiac output is limited by the thickened myocardium through impaired filling and outflow. Mutations in the genes encoding the thick filament components myosin heavy chain and myosin binding protein C (MYH7 and MYBPC3) together explain 75% of inherited HCMs, leading to the observation that HCM is a disease of the sarcomere. Many mutations are “private” or rare variants, often unique to families. In contrast, dilated cardiomyopathy (DCM) is far more genetically heterogeneous, with mutations in genes encoding cytoskeletal, nucleoskeletal, mitochondrial, and calcium-handling proteins. DCM is characterized by enlarged ventricular dimensions and impaired systolic and diastolic function. Private mutations account for most DCMs, with few hotspots or recurring mutations. More than 50 single genes are linked to inherited DCM, including many genes that also link to HCM. Relatively few clinical clues guide the diagnosis of inherited DCM, but emerging evidence supports the use of genetic testing to identify those patients at risk for faster disease progression, congestive heart failure, and arrhythmia. Find the latest version: https://jci.me/62862/pdf Review series Genetic mutations and mechanisms in dilated cardiomyopathy Elizabeth M. McNally, Jessica R. Golbus, and Megan J. Puckelwartz Department of Human Genetics, University of Chicago, Chicago, Illinois, USA. Genetic mutations account for a significant percentage of cardiomyopathies, which are a leading cause of conges- tive heart failure.
    [Show full text]
  • 'Clustering of Nicotinic Acetylcholine Receptors: from the Neuromuscular
    Huh, K H; Fuhrer, C. Clustering of nicotinic acetylcholine receptors: from the neuromuscular junction to interneuronal synapses. Mol. Neurobiol. 2002, 25(1):79-112. Postprint available at: http://www.zora.unizh.ch University of Zurich Posted at the Zurich Open Repository and Archive, University of Zurich. Zurich Open Repository and Archive http://www.zora.unizh.ch Originally published at: Mol. Neurobiol. 2002, 25(1):79-112 Winterthurerstr. 190 CH-8057 Zurich http://www.zora.unizh.ch Year: 2002 Clustering of nicotinic acetylcholine receptors: from the neuromuscular junction to interneuronal synapses Huh, K H; Fuhrer, C Huh, K H; Fuhrer, C. Clustering of nicotinic acetylcholine receptors: from the neuromuscular junction to interneuronal synapses. Mol. Neurobiol. 2002, 25(1):79-112. Postprint available at: http://www.zora.unizh.ch Posted at the Zurich Open Repository and Archive, University of Zurich. http://www.zora.unizh.ch Originally published at: Mol. Neurobiol. 2002, 25(1):79-112 Clustering of nicotinic acetylcholine receptors: from the neuromuscular junction to interneuronal synapses Abstract Fast and accurate synaptic transmission requires high-density accumulation of neurotransmitter receptors in the postsynaptic membrane. During development of the neuromuscular junction, clustering of acetylcholine receptors (AChR) is one of the first signs of postsynaptic specialization and is induced by nerve-released agrin. Recent studies have revealed that different mechanisms regulate assembly vs stabilization of AChR clusters and of the postsynaptic apparatus. MuSK, a receptor tyrosine kinase and component of the agrin receptor, and rapsyn, an AChR-associated anchoring protein, play crucial roles in the postsynaptic assembly. Once formed, AChR clusters and the postsynaptic membrane are stabilized by components of the dystrophin/utrophin glycoprotein complex, some of which also direct aspects of synaptic maturation such as formation of postjunctional folds.
    [Show full text]
  • Profiling of the Muscle-Specific Dystroglycan Interactome Reveals the Role of Hippo Signaling in Muscular Dystrophy and Age-Dependent Muscle Atrophy Andriy S
    Yatsenko et al. BMC Medicine (2020) 18:8 https://doi.org/10.1186/s12916-019-1478-3 RESEARCH ARTICLE Open Access Profiling of the muscle-specific dystroglycan interactome reveals the role of Hippo signaling in muscular dystrophy and age-dependent muscle atrophy Andriy S. Yatsenko1†, Mariya M. Kucherenko2,3,4†, Yuanbin Xie2,5†, Dina Aweida6, Henning Urlaub7,8, Renate J. Scheibe1, Shenhav Cohen6 and Halyna R. Shcherbata1,2* Abstract Background: Dystroglycanopathies are a group of inherited disorders characterized by vast clinical and genetic heterogeneity and caused by abnormal functioning of the ECM receptor dystroglycan (Dg). Remarkably, among many cases of diagnosed dystroglycanopathies, only a small fraction can be linked directly to mutations in Dg or its regulatory enzymes, implying the involvement of other, not-yet-characterized, Dg-regulating factors. To advance disease diagnostics and develop new treatment strategies, new approaches to find dystroglycanopathy-related factors should be considered. The Dg complex is highly evolutionarily conserved; therefore, model genetic organisms provide excellent systems to address this challenge. In particular, Drosophila is amenable to experiments not feasible in any other system, allowing original insights about the functional interactors of the Dg complex. Methods: To identify new players contributing to dystroglycanopathies, we used Drosophila as a genetic muscular dystrophy model. Using mass spectrometry, we searched for muscle-specific Dg interactors. Next, in silico analyses allowed us to determine their association with diseases and pathological conditions in humans. Using immunohistochemical, biochemical, and genetic interaction approaches followed by the detailed analysis of the muscle tissue architecture, we verified Dg interaction with some of the discovered factors.
    [Show full text]
  • Dystrobrevin: Roles in Skeletal Muscle and Its Neuromuscular and Myotendinous Junctions R
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Digital Commons@Becker Washington University School of Medicine Digital Commons@Becker Open Access Publications 2-25-2003 Tyrosine-phosphorylated and nonphosphorylated isoforms of α-dystrobrevin: roles in skeletal muscle and its neuromuscular and myotendinous junctions R. Mark Grady Washington University School of Medicine in St. Louis Mohammed Akaaboune Washington University School of Medicine in St. Louis Alexander L. Cohen Washington University School of Medicine in St. Louis Margaret M. Maimone State University of New York Upstate Medical University Jeff .W Lichtman Washington University School of Medicine in St. Louis See next page for additional authors Follow this and additional works at: http://digitalcommons.wustl.edu/open_access_pubs Part of the Medicine and Health Sciences Commons Recommended Citation Grady, R. Mark; Akaaboune, Mohammed; Cohen, Alexander L.; Maimone, Margaret M.; Lichtman, Jeff .;W and Sanes, Joshua R., ,"Tyrosine-phosphorylated and nonphosphorylated isoforms of α-dystrobrevin: roles in skeletal muscle and its neuromuscular and myotendinous junctions." The ourJ nal of Cell Biology.160,5. 741-752. (2003). http://digitalcommons.wustl.edu/open_access_pubs/625 This Open Access Publication is brought to you for free and open access by Digital Commons@Becker. It has been accepted for inclusion in Open Access Publications by an authorized administrator of Digital Commons@Becker. For more information, please contact [email protected]. Authors R. Mark Grady, Mohammed Akaaboune, Alexander L. Cohen, Margaret M. Maimone, Jeff .W Lichtman, and Joshua R. Sanes This open access publication is available at Digital Commons@Becker: http://digitalcommons.wustl.edu/open_access_pubs/625 Published February 25, 2003 JCBArticle Tyrosine-phosphorylated and nonphosphorylated isoforms of ␣-dystrobrevin: roles in skeletal muscle and its neuromuscular and myotendinous junctions R.
    [Show full text]
  • Neuronal Dystroglycan Is Necessary for Formation and Maintenance of Functional CCK-Positive Basket Cell Terminals on Pyramidal Cells
    10296 • The Journal of Neuroscience, October 5, 2016 • 36(40):10296–10313 Cellular/Molecular Neuronal Dystroglycan Is Necessary for Formation and Maintenance of Functional CCK-Positive Basket Cell Terminals on Pyramidal Cells Simon Fru¨h,1,5 Jennifer Romanos,1,5 Patrizia Panzanelli,2 XDaniela Bu¨rgisser,3 XShiva K. Tyagarajan,1,5 X Kevin P. Campbell,4 XMirko Santello,1,5 and XJean-Marc Fritschy1,5 1Institute of Pharmacology and Toxicology, University of Zurich, 8057 Zurich, Switzerland, 2Department of Neuroscience Rita Levi Montalcini, University of Turin, 10124 Turin, Italy, 3ETH Zurich, 8092 Zurich, Switzerland, 4Howard Hughes Medical Institute, Department of Molecular Physiology and Biophysics, Department of Neurology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa 52242, and 5Neuroscience Center Zurich, University of Zurich and ETH Zurich, 8057 Zurich, Switzerland Distinct types of GABAergic interneurons target different subcellular domains of pyramidal cells, thereby shaping pyramidal cell activity patterns. Whether the presynaptic heterogeneity of GABAergic innervation is mirrored by specific postsynaptic factors is largely unex- plored. Here we show that dystroglycan, a protein responsible for the majority of congenital muscular dystrophies when dysfunctional, has a function at postsynaptic sites restricted to a subset of GABAergic interneurons. Conditional deletion of Dag1, encoding dystrogly- can, in pyramidal cells caused loss of CCK-positive basket cell terminals in hippocampus and neocortex. PV-positive basket cell terminals were unaffected in mutant mice, demonstrating interneuron subtype-specific function of dystroglycan. Loss of dystroglycan in pyrami- dal cells had little influence on clustering of other GABAergic postsynaptic proteins and of glutamatergic synaptic proteins.
    [Show full text]
  • Postmortem Changes in the Myofibrillar and Other Cytoskeletal Proteins in Muscle
    BIOCHEMISTRY - IMPACT ON MEAT TENDERNESS Postmortem Changes in the Myofibrillar and Other C'oskeletal Proteins in Muscle RICHARD M. ROBSON*, ELISABETH HUFF-LONERGAN', FREDERICK C. PARRISH, JR., CHIUNG-YING HO, MARVIN H. STROMER, TED W. HUIATT, ROBERT M. BELLIN and SUZANNE W. SERNETT introduction filaments (titin), and integral Z-line region (a-actinin, Cap Z), as well as proteins of the intermediate filaments (desmin, The cytoskeleton of "typical" vertebrate cells contains paranemin, and synemin), Z-line periphery (filamin) and three protein filament systems, namely the -7-nm diameter costameres underlying the cell membrane (filamin, actin-containing microfilaments, the -1 0-nm diameter in- dystrophin, talin, and vinculin) are listed along with an esti- termediate filaments (IFs), and the -23-nm diameter tubu- mate of their abundance, approximate molecular weights, lin-containing microtubules (Robson, 1989, 1995; Robson and number of subunits per molecule. Because the myofibrils et al., 1991 ).The contractile myofibrils, which are by far the are the overwhelming components of the skeletal muscle cell major components of developed skeletal muscle cells and cytoskeleton, the approximate percentages of the cytoskel- are responsible for most of the desirable qualities of muscle eton listed for the myofibrillar proteins (e.g., myosin, actin, foods (Robson et al., 1981,1984, 1991 1, can be considered tropomyosin, a-actinin, etc.) also would represent their ap- the highly expanded corollary of the microfilament system proximate percentages of total myofibrillar protein. of non-muscle cells. The myofibrils, IFs, cell membrane skel- eton (complex protein-lattice subjacent to the sarcolemma), Some Important Characteristics, Possible and attachment sites connecting these elements will be con- Roles, and Postmortem Changes of Key sidered as comprising the muscle cell cytoskeleton in this Cytoskeletal Proteins review.
    [Show full text]
  • Advanced Fiber Type-Specific Protein Profiles Derived from Adult Murine
    proteomes Article Advanced Fiber Type-Specific Protein Profiles Derived from Adult Murine Skeletal Muscle Britta Eggers 1,2,* , Karin Schork 1,2, Michael Turewicz 1,2 , Katalin Barkovits 1,2 , Martin Eisenacher 1,2, Rolf Schröder 3, Christoph S. Clemen 4,5 and Katrin Marcus 1,2,* 1 Medizinisches Proteom-Center, Medical Faculty, Ruhr-University Bochum, 44801 Bochum, Germany; [email protected] (K.S.); [email protected] (M.T.); [email protected] (K.B.); [email protected] (M.E.) 2 Medical Proteome Analysis, Center for Protein Diagnostics (PRODI), Ruhr-University Bochum, 44801 Bochum, Germany 3 Institute of Neuropathology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, Germany; [email protected] 4 German Aerospace Center, Institute of Aerospace Medicine, 51147 Cologne, Germany; [email protected] 5 Center for Physiology and Pathophysiology, Institute of Vegetative Physiology, Medical Faculty, University of Cologne, 50931 Cologne, Germany * Correspondence: [email protected] (B.E.); [email protected] (K.M.) Abstract: Skeletal muscle is a heterogeneous tissue consisting of blood vessels, connective tissue, and muscle fibers. The last are highly adaptive and can change their molecular composition depending on external and internal factors, such as exercise, age, and disease. Thus, examination of the skeletal muscles at the fiber type level is essential to detect potential alterations. Therefore, we established a protocol in which myosin heavy chain isoform immunolabeled muscle fibers were laser Citation: Eggers, B.; Schork, K.; microdissected and separately investigated by mass spectrometry to develop advanced proteomic Turewicz, M.; Barkovits, K.; profiles of all murine skeletal muscle fiber types.
    [Show full text]
  • B3GALNT2 Is a Gene Associated with Congenital Muscular Dystrophy with Brain Malformations
    European Journal of Human Genetics (2014) 22, 707–710 & 2014 Macmillan Publishers Limited All rights reserved 1018-4813/14 www.nature.com/ejhg SHORT REPORT B3GALNT2 is a gene associated with congenital muscular dystrophy with brain malformations Carola Hedberg*,1, Anders Oldfors1 and Niklas Darin2 Congenital muscular dystrophies associated with brain malformations are a group of disorders frequently associated with aberrant glycosylation of a-dystroglycan. They include disease entities such a Walker–Warburg syndrome, muscle–eye–brain disease and various other clinical phenotypes. Different genes involved in glycosylation of a-dystroglycan are associated with these dystroglycanopathies. We describe a 5-year-old girl with psychomotor retardation, ataxia, spasticity, muscle weakness and increased serum creatine kinase levels. Immunhistochemistry of skeletal muscle revealed reduced glycosylated a-dystroglycan. Magnetic resonance imaging of the brain at 3.5 years of age showed increased T2 signal from supratentorial and infratentorial white matter, a hypoplastic pons and subcortical cerebellar cysts. By whole exome sequencing, the patient was identified to be compound heterozygous for a one-base duplication and a missense mutation in the gene B3GALNT2 (b-1,3-N-acetylgalactos- aminyltransferase 2; B3GalNAc-T2). This patient showed a milder phenotype than previously described patients with mutations in the B3GALNT2 gene. European Journal of Human Genetics (2014) 22, 707–710; doi:10.1038/ejhg.2013.223; published online 2 October 2013 Keywords:
    [Show full text]
  • An Adhesome Comprising Laminin, Dystroglycan and Myosin IIA Is
    © 2014. Published by The Company of Biologists Ltd | Development (2014) 141, 4569-4579 doi:10.1242/dev.116103 RESEARCH ARTICLE An adhesome comprising laminin, dystroglycan and myosin IIA is required during notochord development in Xenopus laevis Nicolas Buisson1, Cathy Sirour2, Nicole Moreau1, Elsa Denker3, Ronan Le Bouffant1, Aline Goullancourt1, Thierry Darriberè 1,* and Valérie Bello1,* ABSTRACT final step, vacuolation, which begins in cells at the anterior and Dystroglycan (Dg) is a transmembrane receptor for laminin that must progesses through to the posterior end of embryos, provides both be expressed at the right time and place in order to be involved stiffness and an increase in cell volume, leading to notochord in notochord morphogenesis. The function of Dg was examined in elongation. Vacuoles are lysosome-related organelles that form Xenopus laevis embryos by knockdown of Dg and overexpression and through a Rab protein and vacuolar-type proton-ATPase-dependent replacement of the endogenous Dg with a mutated form of the protein. acidification (Ellis et al., 2013). The peri-notochordal sheath, in This analysis revealed that Dg is required for correct laminin assembly, combination with the turgor pressure exerted by vacuolated cells, for cell polarization during mediolateral intercalation and for proper provides the rigidity of the notochord that is required for its function differentiation of vacuoles. Using mutations in the cytoplasmic as the major skeletal element of embryos. Although the functions of domain, we identified two sites that are involved in cell polarization integrin in these mechanisms have already been studied (Davidson and are required for mediolateral cell intercalation, and a site that is et al., 2006), there is no information on the involvement of required for vacuolation.
    [Show full text]
  • Α-Dystrobrevin-1 Recruits Α-Catulin to the Α1d- Adrenergic Receptor/Dystrophin-Associated Protein Complex Signalosome
    α-Dystrobrevin-1 recruits α-catulin to the α1D- adrenergic receptor/dystrophin-associated protein complex signalosome John S. Lyssanda, Jennifer L. Whitingb, Kyung-Soon Leea, Ryan Kastla, Jennifer L. Wackera, Michael R. Bruchasa, Mayumi Miyatakea, Lorene K. Langebergb, Charles Chavkina, John D. Scottb, Richard G. Gardnera, Marvin E. Adamsc, and Chris Haguea,1 Departments of aPharmacology and cPhysiology and Biophysics, University of Washington, Seattle, WA 98195; and bDepartment of Pharmacology, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195 Edited by Robert J. Lefkowitz, Duke University Medical Center/Howard Hughes Medical Institute, Durham, NC, and approved October 29, 2010 (received for review July 22, 2010) α1D-Adrenergic receptors (ARs) are key regulators of cardiovascu- pression increases in patients with benign prostatic hypertrophy lar system function that increase blood pressure and promote vas- (12). Through proteomic screening, we discovered that α1D-ARs cular remodeling. Unfortunately, little information exists about are scaffolded to the dystrophin-associated protein complex the signaling pathways used by this important G protein-coupled (DAPC) via the anchoring protein syntrophin (10). Coexpression α “ α receptor (GPCR). We recently discovered that 1D-ARs form a sig- with syntrophins increases 1D-AR plasma membrane expression, nalosome” with multiple members of the dystrophin-associated drug binding, and activation of Gαq/11 signaling after agonist protein complex (DAPC) to become functionally expressed at the activation. Moreover, syntrophin knockout mice lose α1D-AR– plasma membrane and bind ligands. However, the molecular stimulated increases in blood pressure, demonstrating the im- α α mechanism by which the DAPC imparts functionality to the 1D- portance of these essential GIPs for 1D-AR function in vivo (10).
    [Show full text]
  • Critical Review
    IUBMB Life, 61(4): 394–406, April 2009 Critical Review A-kinase Anchoring Proteins: From Protein Complexes to Physiology and Disease Graeme K. Carnegie, Christopher K. Means and John D. Scott Howard Hughes Medical Institute, Department of Pharmacology, University of Washington, School of Medicine, Seattle, Washington, USA or a receptor tyrosine kinase or phosphatase), which results in Summary activation of the receptor or the mobilization of receptor-associ- Protein scaffold complexes are a key mechanism by which a ated proteins to generate some form of intracellular message. common signaling pathway can serve many different functions. There has been a concerted research effort focused to under- Sequestering a signaling enzyme to a specific subcellular envi- stand how the subcellular location of protein kinases and phos- ronment not only ensures that the enzyme is near its relevant targets, but also segregates this activity to prevent indiscrimi- phatases contributes to the regulation of phosphorylation events. nate phosphorylation of other substrates. One family of diverse, Sequestering a signaling enzyme to a specific subcellular envi- well-studied scaffolding proteins are the A-kinase anchoring ronment not only ensures that the enzyme is near to its relevant proteins (AKAPs). These anchoring proteins form multi-protein targets, but also segregates this activity to prevent indiscrimi- complexes that integrate cAMP signaling with other pathways nate phosphorylation of other substrates. Thus protein scaffold and signaling events. In this review, we focus on recent advan- complexes are a key mechanism, by which a common signaling ces in the elucidation of AKAP function. Ó 2009 IUBMB IUBMB Life, 61(4): 394–406, 2009 pathway can serve many different functions.
    [Show full text]
  • What Do We Know About Dystroglycan?
    What Do We Know about Dystroglycan? Daniel Beltran 1 Outline 1. Basement membrane in epithelium and skeletal muscle. 2. Role of dystroglycan. 3. Muscle contraction and repair under normal conditions. 4. Dystroglycanopathies genetics. 5. Dystroglycanopathies muscle pathology. 2 Outline 1. Basement membrane in epithelium and skeletal muscle. 2. Role of dystroglycan. 3. Muscle contraction and repair under normal conditions. 4. Dystroglycanopathies genetics. 5. Dystroglycanopathies muscle pathology. 3 The human body: Cells and connections 4 The human body: Cells and connections Communication and …………… Contact 5 Basement Membrane Basement membrane 6 Basement Membrane Functions Adhesion Mechanic stability Polarity Survival Compartmentalization Proliferation More than just structure 7 Skeletal Muscle 8 Skeletal Muscle Basal Lamina Kjaer (2004). Physiological Reviews, 84: 649. 9 Skeletal Muscle Basal lamina Mouse skeletal muscle Human skeletal muscle 10 Goddeeris et al (2013). Nature, 503:136 Sabatelli et al (2003). Biochim Biophys Acta, 1638:57 Outline 1. Basement membrane in epithelium and skeletal muscle. 2. Role of dystroglycan. 3. Muscle contraction and repair under normal conditions. 4. Dystroglycanopathies genetics. 5. Dystroglycanopathies muscle pathology. 11 Dystroglycan Basement Membrane Laminin α Dystroglycan Membrane β Dystrophin 12 Basement Membrane compaction Colognato 1999. JCB, 145:619 13 Outline 1. Basement membrane in epithelium and skeletal muscle. 2. Role of dystroglycan. 3. Muscle contraction and repair under normal conditions. • Contraction. • Contraction induced muscle damage. • Sarcolemma repair. • Satellite cells mediated muscle repair. • Debris clearance and extracellular remodeling. 14 Muscle Contraction 15 Transmission of force DGC might contribute to “lateral transmission of force” from the sarcomere to the lateral extracellular matrix. 16 Muscle Fiber Repair Han et al (2007).
    [Show full text]