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Functional analysis of titin/connectin N2-B mutations found in cardiomyopathy

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Abstract

Hypertrophic cardiomyopathy and dilated cardiomyopathy are two major clinical phenotypes of “idiopathic” cardiomyopathy. Recent molecular genetic analyses have now revealed that “idiopathic” cardiomyopathy is caused by mutations in genes for sarcomere components. We have recently reported several mutations in titin/connectin gene found in patients with hypertrophic cardiomyopathy or dilated cardiomyopathy. A hypertrophic cardiomyopathy-associated titin/connectin mutation (Arg740Leu) was found to increase the binding to actinin, while other dilated cardiomyopathy-associated titin/connectin mutations (Ala743Val and Val54Met) decreased the binding to actinin and Tcap/telethonin, respectively. We also reported several other mutations in the N2-B region of titin/connectin found in hypertrophic cardiomyopathy and dilated cardiomyopathy. Since the N2-B region expresses only in the heart, it was speculated that functional alterations due to the mutations cause cardiomyopathies. In this study, we investigated the functional changes caused by the N2-B region mutations by using yeast-two-hybrid assays. It was revealed that a hypertrophic cardiomyopathy-associated mutation (Ser3799Tyr) increased the binding to FHL2 protein, whereas a dilated cardiomyopathy-associated mutation (Gln4053ter) decreased the binding. In addition, another TTN mutation (Arg25618Gln) at the is2 region was found in familial DCM. Because FHL2 protein is known to tether metabolic enzymes to N2-B and is2 regions of titin/connectin, these observations suggest that altered recruitment of metabolic enzymes to the sarcomere may play a role in the pathogenesis of cardiomyopathies.

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Abbreviations

DCM:

dilated cardiomyopathy

FHL2:

four and half LIM protein 2

HCM:

hypertrophic cardiomyopathy

MLP:

muscle LIM protein

Tcap:

titin-cap

Y2H:

yeast two hybrid

References

  • Arnber S, Hunter JJ, Ross J Jr., Hongo M, Sansig G, Borg J, Perriard JC, Chien KR, Caroni P, MLP-deficient mice exhibit a disruption of cardiac cytoarchitectural organization, dilated cardiomyopathy, and heart failure Cell 1997;88: 393–403

    Article  PubMed  CAS  Google Scholar 

  • Ashrafian H, Redwood C, Blair E, Watkins H, Hypertrophic cardiomyopathy: a paradigm for myocardial energy depletion Trends Genet 2003;19: 263–268

    Article  PubMed  CAS  Google Scholar 

  • Bang ML, Centner T, Fornoff F, Geach AJ, Gotthardt M, McNabb M, Witt CC, Labeit D, Gregorio CC, Granzier H, Labeit S, The complete gene sequence of titin, expression of an unusual ∼ ∼700-kDa titin isoform, and its interaction with obscuring identify a novel Z-line to I-band linking system Circ Res 2001;89: 1065–1072

    PubMed  CAS  Google Scholar 

  • Bullard, Ferguson C, Minajeva A, Leake MC, Gautel M, Labeit D, Ding L, Labeit S, Horwitz J, Leonard KR, Linke WA, Association of the chaperone alpha B-crystallin with titin in heart muscle J Biol Chem 2003;279: 7917–7924

    Article  PubMed  CAS  Google Scholar 

  • Cazorla O, Wu Y, Irving TC, Granzier H, Titin-based modulation of calcium sensitivity of active tension in mouse skinned cardiac myocytes Circ Res 2001;88: 1028–1035

    PubMed  CAS  Google Scholar 

  • Chan KK, Tsui SK, Lee SM, Luk SC, Liew CC, Fung KP, Waye MM, Lee CY, Molecular cloning and characterization of FHL2, a novel LIM domain protein preferentially expressed in human heart Gene 1998;210: 345–350

    Article  PubMed  CAS  Google Scholar 

  • Chien KR, Genomic circuits and the integrative biology of cardiac diseases Nature 2000;407: 227–232

    Article  PubMed  CAS  Google Scholar 

  • Chien KR, Olson EN, Converging pathways and principles in heart development and disease: CV at CSH Cell 2002;110: 153–162

    Article  PubMed  CAS  Google Scholar 

  • Chu PH, Bardwell WM, Gu Y, Ross Jr. J, Chen J, FHL2 (SLIM3) is not essential for cardiac development and function Mol Cell Biol 2000;20: 7460–7462

    Article  PubMed  CAS  Google Scholar 

  • Genini M, Schwalbe P, Scholl FA, Remppis A, Mattei MG, Schafer BW, Subtractive cloning and characterization of DRAL, a novel LIM-domain protein down-regulated in rhabdomyosarcoma DNA Cell Biol 1997; 16: 433–442

    Article  PubMed  CAS  Google Scholar 

  • Gerull B, Gramlich M, Atherton J, McNabb M, Trombitas K, Sasse-Klaassen S, Seidman JG, Seidman C, Granzier H, Labeit S, Frenneaux M, Thierfelder L, Mutations of TTN, encoding the giant muscle filament titin, cause familial dilated cardiomyopathy Nat Genet 2002;30: 201–204

    Article  PubMed  CAS  Google Scholar 

  • Granzier H, Labeit S, Cardiac titin: an adjustable multi-functional spring J Physiol 2002;541: 335–342

    Article  PubMed  CAS  Google Scholar 

  • Gregorio CC, Trombitas K, Centner T, Kolmerer B, Stier G, Kunke K, Suzuki K, Obermayr F, Herrmann B, Granzier H, Sorimachi H, Labeit S, The NH2 terminus of titin spans the Z-disc: its interaction with a novel 19-kD ligand (T-cap) is required for sarcomeric integrity J Cell Biol 1998;143: 1013–1027

    Article  PubMed  CAS  Google Scholar 

  • Hayashi T, Arimura T, Itoh-Satoh M, Ueda K, Hohda S, Inagaki N, Takahashi M, Hori H, Yasunami M, Nishi H, Koga Y, Nakamura H, Matsuzaki M, Choi BY, Bae SW, You CW, Han KH, Park JE, Knoll R, Hoshijima M, Chien KR, Kimura A, Tcap mutations in hypertrophic cardiomyopathy and dilated cardiomyopathy J Am Coll Cardiol 2004;44: 1992–2001

    Article  CAS  Google Scholar 

  • Itoh-Satoh M, Hayashi T, Nishi H, Koga Y, Arimura T, Koyanagi T, Takahashi M, Hohda S, Ueda K, Nouchi T, Hiroe M, Marumo F, Imaizumi T, Yasunami M, Kimura A, Titin mutations as the molecular basis for dilated cardiomyopathy Biochem Biophys Res Commun 2002;291: 385–393

    Article  PubMed  CAS  Google Scholar 

  • Kamisago M, Sharma SD, DePalma SR, Solomon S, Sharma P, McDonough B, Smoot L, Mullen MP, Woolf PK, Wigle ED, Seidman JG, Seidman CE, Mutations in sarcomere protein genes as a cause of dilated cardiomyopathy N Engl J Med 2000;343: 1688–1696

    Article  PubMed  CAS  Google Scholar 

  • Kimura A, Itoh-Satoh M, Hayashi T, Takahashi M, Arimura T, Yasunami M, Lee SY, Hwang TH, Lee WH, Park JE, (2003). Mutational profiles and molecular pathogenesis of hypertrophic cardiomyopathy and dilated cardiomyopathy in Asian populations In: Dhalla NS, Chockalingam A, Berkovitz HI, Singal PK, (eds). Frontiers in Cardiovascular Health, Kluwer Academic Publishers, Boston pp195–206

    Google Scholar 

  • Knoll R, Hoshijima M, Hoffman HM, Person V, Lorenzen-Schmidt I, Bang ML, Hayashi T, Shiga N, Yasukawa H, Schaper W, McKenna W, Yokoyama M, Schork NJ, Omens JH, McCulloch AD, Kimura A, Gregorio CC, Poller W, Schaper J, Schultheiss HP, Chien KR, The cardiac mechanical stretch sensor machinery involves a Z disc complex that is defective in a subset of human dilated cardiomyopathy Cell 2002;111: 943–955

    Article  PubMed  CAS  Google Scholar 

  • Kong Y, Shelton JM, Rothermel B, Li X, Richardson JA, Bassel-Duby R, Williams RS, Cardiac-specific LIM protein FHL2 modifies the hypertrophic response to beta-adrenergic stimulation Circulation 2001;103: 2731–2738

    PubMed  CAS  Google Scholar 

  • Kumarapeli AR and Wang X, Genetic modification of the heart: chaperones and the cytoskeleton. J Mol Cell Cardiol 2004;37: 1097–1109

    PubMed  CAS  Google Scholar 

  • Labeit S, Kolmerer B, Titins: giant proteins in charge of muscle ultrastructure and elasticity Science 1995; 270: 293–296

    PubMed  CAS  Google Scholar 

  • Lange S, Auerbach D, McLoughlin P, Perriard E, Schafer BW, Perriard JC, Ehler E, Subcellular targeting of metabolic enzymes to titin in heart muscle may be mediated by DRAL/FHL-2 J Cell Sci 2002;115: 4925–4936

    Article  PubMed  CAS  Google Scholar 

  • Li D, Tapscoft T, Gonzalez O, Burch PE, Quinones MA, Zoghbi WA, Hill R, Bachinski LL, Mann DL, Roberts R, Desmin mutation responsible for idiopathic dilated cardiomyopathy Circulation 1999;100: 461–464

    PubMed  CAS  Google Scholar 

  • Li HY, Kotaka M, Kostin S, Lee SM, Kok LD, Chan KK, Tsui SK, Schaper J, Zimmermann R, Lee CY, Fung KP, Waye MM, Translocation of a human focal adhesion LIM-only protein, FHL2, during myofibrinogenesis and identification of LIM2 as the principal determinants of FHL2 focal adhesion localization Cell Motil Cytoskel 2001;48: 11–23

    Article  CAS  Google Scholar 

  • Mohapatra B, Jimenez S, Lin JH, Bowles KR, Coveler KJ, Marx JG, Chrisco MA, Murphy RT, Lurie PR, Schwartz RJ, Elliott PM, Vatta M, McKenna W, Towbin JA, Bowles NE, Mutations in the muscle LIM protein and alpha-actinin-2 genes in dilated cardiomyopathy and endocardial fibroelastosis Mol Genet Metab 2003;80: 207–215

    Article  PubMed  CAS  Google Scholar 

  • Morimoto S, Lu QW, Harada K, Takahashi-Yanaga F, Minakami R, Ohta M, Sasaguri T, Ohtsuki I, Ca (2+)-desensitizing effect of a deletion mutation delta K210 in cardiac troponin T that causes familial cardiomyopathy Proc Natl Acad Sci USA 2002;99: 913–918

    Article  PubMed  CAS  Google Scholar 

  • Mues A, van der Ven PFM, Young P, Furst DO, Gautel M, Two immunoglobulin-like domains of the Z-disc portion of titin interact in a conformation-dependent way with telethonin FEBS Let 1998;428: 111–114

    Article  CAS  Google Scholar 

  • Muller JM, Isele U, Metzger E, Rempel A, Moser M, Pscherer A, Breyer T, Holubarsch C, Buettner R, Schule R, FHL2, a novel tissue-specific coactivator of the androgen receptor EMBO J 2000;19: 359–369

    Article  PubMed  CAS  Google Scholar 

  • Muntoni F, Cau M, Ganau A, Congiu R, Arvedi G, Mateddu A, Marrosu MG, Cianchetti C, Realdi G, Cao A, Melis MA, Brief report: deletion of the dystrophin muscle-promoter region associated with X-linked dilated cardiomyopathy N Engl J Med 1993;329: 921–925

    Article  PubMed  CAS  Google Scholar 

  • Ohtsuka H, Yajima H, Maruyama K, Kimura S, The N-terminal Z repeat 5 of connectin/titin binds to the C-terminal region of alpha-actinin Biochem Biophys Res Commun 1997;235: 1–3

    Article  PubMed  CAS  Google Scholar 

  • Olson TM, Michels VV, Thibodeau SN, Tai YS, Keating MT, Actin mutations in dilated cardiomyopathy, a heritable form of heart failure Science 1998;280: 750–752

    Article  PubMed  CAS  Google Scholar 

  • Pashmforoush M, Pomies P, Peterson KL, Kubalak S, Ross J Jr, Hefti A, Aebi U, Beckerle MC, Chien KR, Adult mice deficient in actinin-associated LIM-domain protein reveal a developmental pathway for right ventricular cardiomyopathy Nat Med 2001;7: 591–597

    Article  PubMed  CAS  Google Scholar 

  • Richardson P, McKenna W, Bristow M, Maisch B, Mautner B, O’Connell J, Olsen E, Thiene G, Goodwin J, Gyarfas I, Martin I, Nordet P, Report of the 1995 World Health Organization/International Society and Federation of Cardiology Task Force on the definition and classification of cardiomyopathies Circulation 1996;93: 841–842

    PubMed  CAS  Google Scholar 

  • Sakamoto A, Ono K, Abe M, Jasmin G, Eki T, Murakami Y, Masaki T, Toyo-oka T, Hanaoka F, Both hypertrophic and dilated cardiomyopathies are caused by mutation of the same gene, delta-sarcoglycan, in hamster: an animal model of disrupted dystrophin-associated glycoprotein complex Proc Natl Acad Sci USA 1997;94: 13873–13878

    Article  PubMed  CAS  Google Scholar 

  • Satoh M, Takahashi M, Sakamoto T, Hiroe M, Marumo F, Kimura A, Structural analysis of the titin gene in hypertrophic cardiomyopathy: identification of a novel disease gene Biochem Biophys Res Commun 1999;262: 411–417

    Article  PubMed  CAS  Google Scholar 

  • Scholl FA, McLoughlin P, Ehler E, de Giovanni C, Schfer BW, DRAL is a p53-responsive gene whose four and a half LIM domain protein product induces apoptosis J Cell Biol 2000;151: 495–505

    Article  PubMed  CAS  Google Scholar 

  • Seidman JG, Seidman C The genetic basis for cardiomyopathy: from mutation identification to mechanistic paradigms Cell 2001;104: 557–567

    Article  PubMed  CAS  Google Scholar 

  • Sorimachi H, Freiburg A, Kolmerer B, Ishiura S, Stier G, Gregorio CC, Labeit D, Linke WA, Suzuki K, Labeit S, Tissue-specific expression and alpha-actinin binding properties of the Z-disc titin: implications for the nature of vertebrate Z-discs J Mol Biol 1997;270: 688–695

    Article  PubMed  CAS  Google Scholar 

  • Tsubata S, Bowles KR, Vatta M, Zintz C, Titus J, Muhonen L, Bowles NE, Towbin JA, Mutations in the human delta-sarcoglycan gene in familial and sporadic dilated cardiomyopathy J Clin Invest 2000;106: 655–662

    Article  PubMed  CAS  Google Scholar 

  • Valle G, Faulkner G, De Antoni A, Pacchioni B, Pallavicini A, Pandolfo D, Tiso N, Toppo S, Trevisan S, Lanfranchi G, Telethonin, a novel sarcomeric protein of heart and skeletal muscle FEBS Let 1997;415: 163–168

    Article  CAS  Google Scholar 

  • Vicart P, Caron A, Guicheney P, Li Z, Prevost MC, Faure A, Chateau D, Chapon F, Tome F, Dupret JM, Paulin D, Fardeau M, A missense mutation in the alpha B-crystallin chaperone gene causes a desmin-related myopathy Nat Genet 1998;20: 92–95

    Article  PubMed  CAS  Google Scholar 

  • Wixler V, Geerts D, Laplantine E, Westhoff D, Smyth N, Aumailley M, Sonnenberg A, Paulsson M, The LIM-only protein DRAL/FHL2 binds to the cytoplasmic domain of several alpha and beta integrin chains and is recruited to adhesion complexes J Biol Chem 2000;275: 33669–33678

    Article  PubMed  CAS  Google Scholar 

  • Xu X, Meiler SE, Zhong TP, Mohideen M, Crossley DA, Burggren WW, Fishman MC, Cardiomyopathy in zebrafish due to mutation in an alternatively spliced exon of titin Nat Genet 2002;30: 205–209

    PubMed  CAS  Google Scholar 

  • Yanaga F, Morimoto S, Ohtsuki I, Ca2+ sensitization and potentiation of the maximum level of myofibrillar ATPase activity caused by mutations of troponin T found in familial hypertrophic cardiomyopathy J Biol Chem 1999;274: 8806–8812

    Article  PubMed  CAS  Google Scholar 

  • Young P, Ferguson C, Banuelos S, Gautel M, Molecular structure of the sarcomeric Z-disk: two types of titin interactions lead to an asymmetrical sorting of alpha-actinin EMBO J 1998;17: 1614–1624

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by Grant-in-Aids for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan and a research grant from the Ministry of Health, Labor and Welfare, Japan and Mitsui Life Social Welfare Foundation. We are grateful to Drs H. Toshima, H. Nishi, Y. Koga, K. Matsuyama, T. Sakamoto, K. Kawai, K.␣Kawamura, R. Kusukawa, M. Nagano, Y.␣Nimura, R. Okada, T. Sugimoto, H. Tanaka, H. Yasuda, K. Fukuda, S. Ogawa, A. Matsumori, S. Sasayama, R. Nagai, and Y. Yazaki for their contributions in clinical evaluation and blood sampling from patients with HCM and DCM

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Authors and Affiliations

  1. Department of Molecular Pathogenesis, Medical Research Institute, Tokyo Medical and Dental University, 2-3-10 Kandasurugadai, Chiyoda-Ku, 101-0062, Tokyo , Japan

    YUJI MATSUMOTO, TAKEHARU HAYASHI, NATSUKO INAGAKI, MEGUMI TAKAHASHI, SHITOSHI HIROI, TAKEYUKI NAKAMURA, TAKURO ARIMURA, MICHIO YASUNAMI &  AKINORI KIMURA

  2. Division of Cardiovascular Medicine, Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, Nagasaki, Japan

    YUJI MATSUMOTO, NAOTO ASHIZAWA & KATSUSUKE YANO

  3. Department of Cardiovascular Medicine, Okayama University School of Medicine, Okayama, Japan

    KAZUFUMI NAKAMURA & TORU OHE

  4. Laboratory of Genome Diversity, School of Biomedical Science, Tokyo Medical and Dental University, Tokyo, Japan

    MICHIO YASUNAMI &  AKINORI KIMURA

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  1. YUJI MATSUMOTO
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  8. KAZUFUMI NAKAMURA
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  12. KATSUSUKE YANO
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  13. AKINORI KIMURA
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Correspondence to AKINORI KIMURA.

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MATSUMOTO, Y., HAYASHI, T., INAGAKI, N. et al. Functional analysis of titin/connectin N2-B mutations found in cardiomyopathy. J Muscle Res Cell Motil 26, 367–374 (2005). https://doi.org/10.1007/s10974-005-9018-5

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