DOCSLIB.ORG

Viewed in [38])

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Viewed in [38]) Molecular Cancer BioMed Central Research Open Access The parafibromin tumor suppressor protein interacts with actin-binding proteins actinin-2 and actinin-3 Sunita K Agarwal*, William F Simonds and Stephen J Marx Address: National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, Maryland, USA Email: Sunita K Agarwal* - [email protected]; William F Simonds - [email protected]; Stephen J Marx - [email protected] * Corresponding author Published: 7 August 2008 Received: 24 April 2008 Accepted: 7 August 2008 Molecular Cancer 2008, 7:65 doi:10.1186/1476-4598-7-65 This article is available from: http://www.molecular-cancer.com/content/7/1/65 © 2008 Agarwal et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Germline and somatic inactivating mutations in the HRPT2 gene occur in the inherited hyperparathyroidism-jaw tumor syndrome, in some cases of parathyroid cancer and in some cases of familial hyperparathyroidism. HRPT2 encodes parafibromin. To identify parafibromin interacting proteins we used the yeast two-hybrid system for screening a heart cDNA library with parafibromin as the bait. Results: Fourteen parafibromin interaction positive preys representing 10 independent clones encoding actinin-2 were isolated. Parafibromin interacted with muscle alpha-actinins (actinin-2 and actinin-3), but not with non-muscle alpha-actinins (actinin-1 and actinin-4). The parafibromin-actinin interaction was verified by yeast two-hybrid, GST pull-down, and co-immunoprecipitation. Yeast two-hybrid analysis revealed that the N-terminal region of parafibromin interacted with actinins. In actin sedimentation assays parafibromin did not dissociate skeletal muscle actinins from actin filaments, but interestingly, parafibromin could also bundle/cross-link actin filaments. Parafibromin was predominantly nuclear in undifferentiated proliferating myoblasts (C2C12 cells), but in differentiated C2C12 myotubes parafibromin co-localized with actinins in the cytoplasmic compartment. Conclusion: These data support a possible contribution of parafibromin outside the nucleus through its interaction with actinins and actin bundling/cross-linking. These data also suggest that actinins (and actin) participate in sequestering parafibromin in the cytoplasmic compartment. Background somatic inactivation of HRPT2 is common in parathyroid Parafibromin is encoded by the HRPT2 gene located on tumor of HPT-JT and in sporadic parathyroid cancer [5-7]. chromosome 1q31[1] (OMIM# 145001 and 607393). HRPT2 inactivating mutations in the germline occur in Parafibromin has three possible nuclear localization sig- the hyperparathyroidism-jaw tumor (HPT-JT) syndrome, nals; it is predominantly found in the nucleus [8-16] and characterized by hyperparathyroidism (parathyroid ade- in the nucleolus [17]. The C-terminal part of parafibromin noma or carcinoma) (90%), fibrous-ossifying jaw tumors (ORF = 531 amino acids) is homologous to yeast Cdc73p (30%), and bilateral renal cysts (10%) [1,2]. Germline (ORF = 393 amino acids), an RNA polymerase II accessory HRPT2 mutations have also been detected in some cases protein [18]. Analogous to the presence of yeast Cdc73p of familial hyperparathyroidism [1,3,4], and biallelic in the yeast Paf1 protein complex, parafibromin was Page 1 of 11 (page number not for citation purposes) Molecular Cancer 2008, 7:65 http://www.molecular-cancer.com/content/7/1/65 found in the human and Drosophila PAF1 protein complex were also positive for interaction with GalDBD-parafibro- [10,19-21]. Genetic interaction studies in Drosophila and min (Table 2). Actinin-2 and FHL2 each belong to fami- protein interaction studies in mammalian cells have lies of proteins; therefore, independent members of each shown an interaction of parafibromin with β-catenin, family were tested for interaction with parafibromin mediating a functional association between Wnt signaling (Table 2). Parafibromin selectively interacted with the and the PAF1 complex [22]. skeletal muscle actinins (actinin-2 and actinin-3), but did not interact with the non-muscle actinins (actinin-1 and The yeast two-hybrid system has been extensively used for actinin-4). Among the four FHL family members, parafi- the identification and analysis of protein-protein interac- bromin only interacted with FHL2. tions [23,24]. To isolate parafibromin interacting proteins we conducted a yeast two-hybrid library screen, and we GST pull-down and co-immunoprecipitation of discovered an interaction between parafibromin and mus- parafibromin interacting proteins cle actinins. To verify the in vivo finding in yeast of parafibromin inter- action with actinins, desmin and FHL2, GST pull-down Results assays were performed (Figure 1). Protein extracts from Parafibromin interacting proteins HEK293 cells expressing parafibromin-myc-his were To identify proteins that interact with parafibromin, a tested for binding to glutathione sepharose beads coupled yeast two-hybrid screen was performed with parafibromin with GST alone, or with GST-actinin-2, GST-actinin-3, as the bait. Because no parafibromin interacting proteins GST-desmin, or GST-FHL2. Parafibromin was specifically had been obtained by screening a human brain library associated with GST-actinin-2, GST-actinin-3, but not (data not shown) and because of high-level expression of with GST alone, GST-desmin, or GST-FHL2 (Figure 1A parafibromin in heart tissue [1], a human heart cDNA and Figure 1B). Desmin and FHL2 were not analyzed fur- library was screened. From 106 yeast transformants, 76 ther. colonies grew in selection medium; 36 of 76 were positive for the second reporter LacZ; upon retransformation 27 of The interaction of parafibromin with actinins was further 36 exhibited interaction with parafibromin. Sequencing characterized in vivo by co-immunoprecipitation of of the inserts from the 27 plasmids revealed cDNAs actinin-2 or actinin-3 from protein extracts of HEK293 encoding 6 different proteins (Table 1). There were 14 cells transfected with parafibromin-myc-his or menin- ACTN2 (actinin-2) sequences representing 10 independ- myc-his together with FLAG-tagged actinin-2 or actinin-3. ent clones, 5 FHL2 (four and a half LIM-domains 2) FLAG-actinin-2 and FLAG-actinin-3 co-precipitated with sequences representing 3 independent clones, 3 DES parafibromin-myc-his but not with menin-myc-his (Fig- (desmin) sequences representing 3 independent clones, ure 1C). and one sequence each of DMPK (dystrophia myotonica protein kinase), CLTCL1 (clathrin heavy chain like-1) and Interacting regions of parafibromin and actinin EEA1 (early endosome antigen 1). All sequences were in- N- or C-terminal deletion constructs of parafibromin or frame with the Gal4AD. actinin-3 were tested for interactions, in yeast two-hybrid assays (Figure 2). All three C-terminal truncations of Since actinin-2, FHL2, and desmin were represented by parafibromin interacted with actinin-3, whereas the two multiple independent library clones, they were analyzed N-terminal truncations of parafibromin did not interact further for interaction of their full-length coding regions with actinin-3. Each of the 10 yeast two-hybrid library with parafibromin. Full-length coding regions of all three clones (actinin-2) that was isolated as a parafibromin Table 1: Yeast Two-hybrid Library Screening Gene Entrez Description Number Number of Name Gene ID of Independent Colonies Clones* ACTN2 88 Skeletal muscle actinin, alpha-2 (actinin-2) 14 10 FHL2 2274 Four and a half LIM-domains 2 5 3 DES 1674 Desmin 3 3 DMPK 1760 Dystrophia myotonica protein kinase 3 1 CLTCL1 8218 Clathrin heavy chain like-1 1 1 EEA1 8411 Early endosome antigen 1 1 1 *ACTN2: 10 independent clones starting at amino acid (aa) 323, 327, 329, 333, 340, 360, 370, 377, 381 and 390 to the stop codon after aa 894; FHL2: 3 independent full-length clones differed in the length of the 5' UTR (63 bp, 75 bp, and 150 bp); DES: 3 independent clones starting after aa 142, 115, and 203 to the stop codon after aa 468. Page 2 of 11 (page number not for citation purposes) Molecular Cancer 2008, 7:65 http://www.molecular-cancer.com/content/7/1/65 Table 2: Yeast Two-hybrid Interaction† of Parafibromin Fused to Gal4DBD with Full-length Proteins Fused to Gal4AD Activation Domain Fusion β-Galactosidase Assay 1 Actinin-1 - 2 Actinin-2* ++ 3 Actinin-3* +++ 4 Actinin-4 - 5 α-actin - 6FHL-1 - 7FHL-2 ++ 8FHL-3 - 9FHL-4 - 10 Desmin ++ 11 β-catenin* + †Yeast strain MaV203 that contains the Gal4-LacZ reporter or EGY48/pSH-18 (for β-catenin) that contains the LexA-LacZ reporter was transformed with plasmids expressing Gal4DBD- or LexA-DBD- fusion proteins together with plasmids expressing Gal4AD-fusion proteins. Transformed yeast colonies selected in SD medium lacking Leu and Trp (Leu, Trp and Ura for EGY48/pSH-18) were scored for β-galactosidase activity in colony filter lift assays. Interaction is indicated by β-galactosidase activity level. -, +, ++, or +++ indicate no, low, medium or high levels of β-galactosidase activity, respectively. None of the Gal4AD fusions interacted with Gal4DBD alone or with Gal4DBD-p53 (aa 72–390 of mouse p53). The Gal4DBD-parafibromin did not interact with Gal4AD alone or Gal4AD-TAg (aa 87–708 of SV-40 T-antigen). *Interaction was also positive with parafibromin fused to LexA-DBD (construct from [22]). interactor was N-terminally truncated at amino acids 323, VerificationwithFigure actinin 1 of yeast two-hybrid interaction of parafibromin 327, 329, 333, 340, 360, 370, 377, 381 and 390 (Table 1). Verification of yeast two-hybrid interaction of parafi- The N-terminus was located in the first spectrin repeat, bromin with actinin. (A) GST or GST-fusion proteins used and each interacted strongly with parafibromin. Further for in vitro binding assay were analyzed by SDS-PAGE and N-terminal truncations, deleting spectrin repeat 2 and 3 of stained with Coomassie Blue.
Load more

Recommended publications
  • Desmin Interacts Directly with Mitochondria
    International Journal of Molecular Sciences Article Desmin Interacts Directly with Mitochondria Alexander A. Dayal 1, Natalia V. Medvedeva 1, Tatiana M. Nekrasova 1, Sergey D. Duhalin 1, Alexey K. Surin 1,2 and Alexander A. Minin 1,* 1 Institute of Protein Research of Russian Academy of Sciences, Vavilova st., 34, 119334 Moscow, Russia; [email protected] (A.A.D.); [email protected] (N.V.M.); [email protected] (T.M.N.); [email protected] (S.D.D.); [email protected] (A.K.S.) 2 Pushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Prospekt Nauki 6, Pushchino, 142290 Moscow Region, Russia * Correspondence: [email protected] Received: 14 October 2020; Accepted: 26 October 2020; Published: 30 October 2020 Abstract: Desmin intermediate filaments (IFs) play an important role in maintaining the structural and functional integrity of muscle cells. They connect contractile myofibrils to plasma membrane, nuclei, and mitochondria. Disturbance of their network due to desmin mutations or deficiency leads to an infringement of myofibril organization and to a deterioration of mitochondrial distribution, morphology, and functions. The nature of the interaction of desmin IFs with mitochondria is not clear. To elucidate the possibility that desmin can directly bind to mitochondria, we have undertaken the study of their interaction in vitro. Using desmin mutant Des(Y122L) that forms unit-length filaments (ULFs) but is incapable of forming long filaments and, therefore, could be effectively separated from mitochondria by centrifugation through sucrose gradient, we probed the interaction of recombinant human desmin with mitochondria isolated from rat liver. Our data show that desmin can directly bind to mitochondria, and this binding depends on its N-terminal domain.
    [Show full text]
  • Contralateral Recurrence of Aggressive Fibromatosis in a Young Woman: a Case Report and Review of the Literature
    ONCOLOGY LETTERS 10: 325-328, 2015 Contralateral recurrence of aggressive fibromatosis in a young woman: A case report and review of the literature CHRISTOPHER J. SCHMOYER, HARMAR D. BRERETON and ERIC W. BLOMAIN Clinical Faculty, Department of Medicine, The Commonwealth Medical College, Scranton, PA 18509, USA Received August 9, 2014; Accepted April 24, 2015 DOI: 10.3892/ol.2015.3215 Abstract. Aggressive fibromatosis (AF) is a benign and shoulder girdle. Individuals with familial adenomatous non-encapsulated tumor of mesenchymal origin, with a polyposis (FAP) or Gardner's syndrome have a 1,000 times tendency for local spread along fascial planes. Local inva- greater risk for developing the disease due to inheritance of sion can lead to extensive morbidity and even mortality due the adenomatous polyposis coli (APC) gene (3). These patients to destruction of the bones, organs and soft tissues. This rare may present with intra-abdominal lesions following colonic lesion is observed 1,000 times more frequently in patients with resection (4). While AF does not metastasize, local recurrence familial adenomatous polyposis or Gardner's syndrome due to is common. Distant recurrence is extremely rare, but is typi- the inheritance of the adenomatous polyposis coli (APC) gene. cally observed in those with a new primary tumor associated While AF does not metastasize, local recurrence is common. with the APC mutation. The present study reports the case of Distant recurrence is extremely rare, but is observed in those a 20-year-old female with sporadic contralateral recurrence of with a germ line APC mutation. The present study details clinically diagnosed AF and no familial predisposition.
    [Show full text]
  • Anti-ACTN2 Monoclonal Antibody, Clone FQ3630Z (DCABH-9438) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use
    Anti-ACTN2 monoclonal antibody, clone FQ3630Z (DCABH-9438) This product is for research use only and is not intended for diagnostic use. PRODUCT INFORMATION Product Overview Rabbit monoclonal to Sarcomeric Alpha Actinin Antigen Description F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein. Immunogen A synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to residues on the N-terminus of human Sarcomeric Alpha Actinin Isotype IgG Source/Host Rabbit Species Reactivity Mouse, Rat, Human Clone FQ3630Z Purity Tissue culture supernatant Conjugate Unconjugated Applications WB, IP, IHC-P Positive Control Human skeletal muscle, rat heart and human heart lysates and human muscle tissue. Format Liquid Size 100 μl Buffer pH: 7.40; Preservative: 0.01% Sodium azide; Constituents: 50% Glycerol, 0.05% BSA Preservative 0.01% Sodium Azide Storage Store at -20°C. Stable for 12 months at -20°C GENE INFORMATION Gene Name ACTN2 actinin, alpha 2 [ Homo sapiens ] Official Symbol ACTN2 45-1 Ramsey Road, Shirley, NY 11967, USA Email: [email protected] Tel: 1-631-624-4882 Fax: 1-631-938-8221 1 © Creative Diagnostics All Rights Reserved Synonyms ACTN2; actinin, alpha 2; alpha-actinin-2; F-actin cross-linking protein; alpha-actinin skeletal muscle; CMD1AA; Entrez Gene ID 88 Protein Refseq NP_001094 UniProt ID P35609 Chromosome Location 1q42-q43 Pathway Activation of NMDA receptor upon glutamate binding and postsynaptic events,
    [Show full text]
  • Differential Organization of Desmin and Vimentin in Muscle Is Due to Differences in Their Head Domains Robert B
    Differential Organization of Desmin and Vimentin in Muscle Is Due to Differences in Their Head Domains Robert B. Cary and Michael W. Klymkowsky Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347 Abstract. In most myogenic systems, synthesis of the aggregates. In embryonic epithelial cells, both vimen- intermediate filament (IF) protein vimentin precedes tin and desmin formed extended IF networks. Vimen- the synthesis of the muscle-specific IF protein desmin. tin and desmin differ most dramatically in their NH:- In the dorsal myotome of the Xenopus embryo, how- terminal "head" regions. To determine whether the ever, there is no preexisting vimentin filament system head region was responsible for the differences in the and desmin's initial organization is quite different from behavior of these two proteins, we constructed plas- that seen in vimentin-containing myocytes (Cary and mids encoding chimeric proteins in which the head of Klymkowsky, 1994. Differentiation. In press.). To de- one was attached to the body of the other. In muscle, termine whether the organization of IFs in the Xeno- the vimentin head-desmin body (VDD) polypeptide pus myotome reflects features unique to Xenopus or is formed longitudinal IFs and massive IF bundles like due to specific properties of desmin, we used the in- vimentin. The desmin head-vimentin body (DVV) jection of plasmid DNA to drive the synthesis of polypeptide, on the other hand, formed IF meshworks vimentin or desmin in myotomal cells. At low levels and non-filamentous structures like desmin. In em- of accumulation, exogenous vimentin and desmin both bryonic epithelial cells DVV formed a discrete fila- enter into the endogenous desmin system of the myo- ment network while VDD did not.
    [Show full text]
  • Genetic Cardiomyopathies: the Lesson Learned from Hipscs
    Journal of Clinical Medicine Review Genetic Cardiomyopathies: The Lesson Learned from hiPSCs Ilaria My 1,2 and Elisa Di Pasquale 2,3,* 1 Department of Biomedical Sciences, Humanitas University, Pieve Emanuele, 20090 Milan, Italy; [email protected] 2 Humanitas Clinical and Research Center—IRCCS, Rozzano, 20089 Milan, Italy 3 Institute of Genetic and Biomedical Research (IRGB)—UOS of Milan, National Research Council (CNR), 20138 Milan, Italy * Correspondence: [email protected] Abstract: Genetic cardiomyopathies represent a wide spectrum of inherited diseases and constitute an important cause of morbidity and mortality among young people, which can manifest with heart failure, arrhythmias, and/or sudden cardiac death. Multiple underlying genetic variants and molecular pathways have been discovered in recent years; however, assessing the pathogenicity of new variants often needs in-depth characterization in order to ascertain a causal role in the disease. The application of human induced pluripotent stem cells has greatly helped to advance our knowledge in this field and enabled to obtain numerous in vitro patient-specific cellular models useful to study the underlying molecular mechanisms and test new therapeutic strategies. A milestone in the research of genetically determined heart disease was the introduction of genomic technologies that provided unparalleled opportunities to explore the genetic architecture of cardiomyopathies, thanks to the generation of isogenic pairs. The aim of this review is to provide an overview of the main research that helped elucidate the pathophysiology of the most common genetic cardiomyopathies: hypertrophic, dilated, arrhythmogenic, and left ventricular noncompaction cardiomyopathies. A special focus is provided on the application of gene-editing techniques in understanding key disease characteristics and on the therapeutic approaches that have been tested.
    [Show full text]
  • Identification of Previously Unrecognized FAP in Children With
    European Journal of Human Genetics (2015) 23, 715–718 & 2015 Macmillan Publishers Limited All rights reserved 1018-4813/15 www.nature.com/ejhg SHORT REPORT Identification of previously unrecognized FAP in children with Gardner fibroma Joana Vieira1,5, Carla Pinto1,5, Mariana Afonso2,5, Maria do Bom Sucesso3, Paula Lopes2, Manuela Pinheiro1, Isabel Veiga1, Rui Henrique2,4 and Manuel R Teixeira*,1,4 Fibromatous soft tissue lesions, namely desmoid-type fibromatosis and Gardner fibroma, may occur sporadically or as a result of inherited predisposition (as part of familial adenomatous polyposis, FAP). Whereas desmoid-type fibromatosis often present b-catenin overexpression (by activating CTNNB1 somatic variants or APC biallelic inactivation), the pathogenetic mechanisms in Gardner fibroma are unknown. We characterized in detail Gardner fibromas diagnosed in two infants to evaluate their role as sentinel lesions of previously unrecognized FAP. In the first infant we found a 5q deletion including APC in the tumor and the novel APC variant c.4687dup in constitutional DNA. In the second infant we found the c.5826_5829del and c.1678A4T APC variants in constitutional and tumor DNA, respectively. None of the constitutional APC variants occurred de novo and both tumors showed nuclear staining for b-catenin and no CTNNB1 variants. We present the first comprehensive characterization of the pathogenetic mechanisms of Gardner fibroma, which may be a sentinel lesion of previously unrecognized FAP families. European Journal of Human Genetics (2015) 23, 715–718; doi:10.1038/ejhg.2014.144; published online 30 July 2014 INTRODUCTION MATERIALS AND METHODS Familial adenomatous polyposis (FAP) is an autosomal dominant The first case was a 5-month-old child who had two lumbar subcutaneous disease caused by APC constitutional variants.
    [Show full text]
  • Disease-Proportional Proteasomal Degradation of Missense Dystrophins
    Disease-proportional proteasomal degradation of missense dystrophins Dana M. Talsness, Joseph J. Belanto, and James M. Ervasti1 Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota–Twin Cities, Minneapolis, MN 55455 Edited by Louis M. Kunkel, Children’s Hospital Boston, Harvard Medical School, Boston, MA, and approved September 1, 2015 (received for review May 5, 2015) The 427-kDa protein dystrophin is expressed in striated muscle insertions or deletions (indels) represent ∼7% of the total DMD/ where it physically links the interior of muscle fibers to the BMD population (13). When indel mutations cause a frameshift, they extracellular matrix. A range of mutations in the DMD gene encod- can specifically be targeted by current exon-skipping strategies (15). ing dystrophin lead to a severe muscular dystrophy known as Du- Patients with missense mutations account for only a small percentage chenne (DMD) or a typically milder form known as Becker (BMD). of dystrophinopathies (<1%) (13), yet they represent an orphaned Patients with nonsense mutations in dystrophin are specifically tar- subpopulation with an undetermined pathomechanism and no cur- geted by stop codon read-through drugs, whereas out-of-frame de- rent personalized therapies. letions and insertions are targeted by exon-skipping therapies. Both The first missense mutation reported to cause DMD was L54R treatment strategies are currently in clinical trials. Dystrophin mis- in ABD1 of an 8-y-old patient (16). Another group later reported sense mutations, however, cause a wide range of phenotypic se- L172H, a missense mutation in a structurally analogous location of verity in patients. The molecular and cellular consequences of such ABD1 (17), yet this patient presented with mild symptoms at 42 mutations are not well understood, and there are no therapies spe- years of age.
    [Show full text]
  • Final Draft Masterthesis Gabbin
    MASTER THESIS Thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Engineering at the University of Applied Sciences Technikum Wien Degree Program in Tissue Engineering and Regenerative Medicine Modeling of a Novel Filamin-C Mutation in Restrictive Cardiomyopathy using hiPSC- derived Cardiomyocytes By: Beatrice Gabbin, BSc Student Number: 1810692024 Supervisor 1: Dr. William Pu, MD Dr. Maksymilian Prondzynski, PhD Supervisor 2: Dorota Szwarc, MSc Boston, September 29, 2020 Declaration of Authenticity “As author and creator of this work to hand, I confirm with my signature knowledge of the relevant copyright regulations governed by higher education acts (see Urheberrechtsgesetz/ Austrian copyright law as amended as well as the Statute on Studies Act Provisions / Examination Regulations of the UAS Technikum Wien as amended). I hereby declare that I completed the present work independently and that any ideas, whether written by others or by myself, have been fully sourced and referenced. I am aware of any consequences I may face on the part of the degree program director if there should be evidence of missing autonomy and independence or evidence of any intent to fraudulently achieve a pass mark for this work (see Statute on Studies Act Provisions / Examination Regulations of the UAS Technikum Wien as amended). I further declare that up to this date I have not published the work to hand nor have I presented it to another examination board in the same or similar form. I affirm that the version submitted matches the version in the upload tool.” Boston, September 29, 2020 Place, Date Signature Abstract The purpose of this project was to establish an in vitro model for a novel FLNC missense mutation (c.7416_7418delGAA/p.Glu2472_Asn2473delAsp), which was identified at the Boston Children’s Hospital in a patient diagnosed with restrictive cardiomyopathy (RCM).
    [Show full text]
  • IL-33 Activates Tumor Stroma to Promote Intestinal Polyposis
    IL-33 activates tumor stroma to promote PNAS PLUS intestinal polyposis Rebecca L. Maywalda, Stephanie K. Doernerb, Luca Pastorellic,d,e, Carlo De Salvoc, Susan M. Bentona, Emily P. Dawsona, Denise G. Lanzaa, Nathan A. Bergerb,f,g,h, Sanford D. Markowitzh,i,j, Heinz-Josef Lenzk, Joseph H. Nadeaul, Theresa T. Pizarroc, and Jason D. Heaneya,m,n,1 aDepartment of Molecular and Human Genetics, mDan L. Duncan Cancer Center, and nTexas Medical Center Digestive Diseases Center, Baylor College of Medicine, Houston, TX 77030; Departments of bGenetics and Genome Sciences, cPathology, fBiochemistry, and iMedicine, gCenter for Science, Health and Society, hCase Comprehensive Cancer Center, and jUniversity Hospitals Case Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH 44106; dDepartment of Biomedical Sciences for Health, University of Milan, Milan 20133, Italy; eGastroenterology and Gastrointestinal Endoscopy Unit, Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Donato, San Donato Milanese 20097, Italy; lPacific Northwest Research Institute, Seattle, WA 98122; and kDivision of Medical Oncology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033 Edited by Charles A. Dinarello, University of Colorado Denver, Aurora, CO, and approved April 2, 2015 (received for review November 25, 2014) Tumor epithelial cells develop within a microenvironment consist- epithelial restitution and proliferation (6, 7). The CRC stroma ing of extracellular matrix, growth factors, and cytokines produced acquires a similar activated phenotype and produces the same by nonepithelial stromal cells. In response to paracrine signals from soluble factors and ECM components associated with inflammation tumor epithelia, stromal cells modify the microenvironment to pro- and wound healing to promote proliferation and survival of trans- mote tumor growth and metastasis.
    [Show full text]
  • Desmin and Vimentin
    Proc. NatL Acad. Sci. USA Vol. 78, No. 7, pp. 4120-4123, July 1981 Biochemistry Comparison of the proteins of two immunologically distinct intermediate-sized filaments by amino acid sequence analysis: Desmin and vimentin (cytoskeleton/differentiation/eye lens/keratin/tropomyosin) NORBERT GEISLER AND KLAus WEBER Max Planck Institute for Biophysical Chemistry, D-3400 Goettingen, Federal Republic of Germany Communicated by Manfred Eigen, March 25, 1981 ABSTRACT Although all intermediate-sized filaments (10-nm In spite of the differences revealed by two-dimensional gel filaments) seem to show similar morphology and share a number analysis and the lack of immunological crossreactivity experi- of biochemical properties, different cell- and tissue-specific sub- enced with many antibodies, the structural proteins of these classes have been distinguished by immunological experiments filaments share several distinct properties. The filaments show and by differences in apparent molecular weights and isoelectric a very similar ultrastructure and morphology and belong to the points of the major constituent proteins. In order to understand highly helical k-m-e-f-type ofproteins (see, for instance, ref. 7). the degree ofpossible homology between these proteins, we have In addition they are all insoluble in physiological buffers (for begun amino acid sequence analysis ofthe polypeptides. Here we references see above) and, with the exception of certain neu- characterize a large fragment ofchicken gizzard and pig stomach molecular weights desmin as well as the corresponding fragment from porcine eye rofilament proteins, contain polypeptides of lens vimentin. The fragments are situated at the carboxyl end and in the range 45,000-68,000 (for references see refs.
    [Show full text]
  • Desmin Is a Modifier of Dystrophic Muscle Features in Mdx Mice
    bioRxiv preprint doi: https://doi.org/10.1101/742858; this version posted August 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Desmin is a modifier of dystrophic muscle features in Mdx mice Arnaud Ferry (1,2), Julien Messéant (1), Ara Parlakian (3), Mégane Lemaitre (1), Pauline Roy (1), Clément Delacroix (1), Alain Lilienbaum (4), Yeranuhi Hovhannisyan (3), Denis Furling (1), Arnaud Klein (1), Zhenlin Li (3), Onnik Agbulut (3) 1-Sorbonne Université, INSERM U974, CNRS UMR7215, Institut de Myologie, Paris, F- 75013 France 2-Université de Paris, Institut des Sciences du Sport Santé de Paris, Paris, F-75006 France 3- Sorbonne Université, Institut de Biologie Paris-Seine (IBPS), UMR CNRS 8256, INSERM ERL U1164, Biological Adaptation and Ageing, Paris, F-75005 France. 4-Université de Paris, Unité de Biologie Fonctionnelle et Adaptative, CNRS UMR 8251, Paris, F-75013 France. Corresponding author: Arnaud Ferry 1 bioRxiv preprint doi: https://doi.org/10.1101/742858; this version posted August 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract (175 words) Duchenne muscular dystrophy (DMD) is a severe neuromuscular disease, caused by dystrophin deficiency. Desmin is like dystrophin associated to costameric structures bridging sarcomeres to extracellular matrix that are involved in force transmission and skeletal muscle integrity. In the present study, we wanted to gain further insight into the roles of desmin which expression is increased in the muscle from the mouse Mdx DMD model.
    [Show full text]
  • Molecular Interactions of the Mammalian Intermediate Filament Protein Synemin with Cytoskeletal Proteins Present in Adhesion Sites Ning Sun Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2008 Molecular interactions of the mammalian intermediate filament protein synemin with cytoskeletal proteins present in adhesion sites Ning Sun Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Molecular Biology Commons Recommended Citation Sun, Ning, "Molecular interactions of the mammalian intermediate filament protein synemin with cytoskeletal proteins present in adhesion sites" (2008). Retrospective Theses and Dissertations. 15814. https://lib.dr.iastate.edu/rtd/15814 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Molecular interactions of the mammalian intermediate filament protein synemin with cytoskeletal proteins present in adhesion sites by Ning Sun A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Molecular, Cellular, and Developmental Biology Program of Study Committee Richard M. Robson, Major Professor Ted W. Huiatt Steven M. Lonergan Jo Anne Powell-Coffman Linda Ambrosio Iowa State University Ames, Iowa 2008 Copyright © Ning Sun, 2008. All rights reserved. 3316170
    [Show full text]