Signalling-Pathways Regulating the Dynamic Localization of Z-disc-Associated Proteins
Final Report Abstract
The Z-disc is a major structural part of the myofibril which has to withstand enormous mechanical strain. It is composed of a limited number of structural proteins but a plethora of polypeptides are associated with the Z-disc, at least transiently. This dynamic nature is important to allow for continuous replacement of damaged components and to keep up adaptability to varying physiological needs. The main aim of our project was to decipher how this dynamic behaviour of Z-disc components is regulated, and in particular, which signalling pathways influence protein interactions and protein turnover. Extensive biochemical studies allowed us to establish the most comprehensive map of proteinprotein interactions at the Z-disc known to date. To understand how these interactions are regulated, we established in close collaboration with P4 a map of protein phosphorylations elicited by contractile activity. This data provided a basis for detailed analyses of protein interaction dynamics, which we investigated at the sub-cellular level using BiFC and FRAP. We focused our investigations on the large actin-binding protein filamin C (FLNc) and its ligands and found that they are particularly dynamic Z-disc-associated proteins and that phosphorylation fine-tunes this dynamic behavior. The main function of these proteins seems to sense mechanically damaged areas in myofibrils and to initiate subsequent repair. In this context we found, in close collaboration with P10, that the homeostasis of FLNc is controlled by interactions with HSPB8, BAG3 (linking to chaperoneassisted selective autophagy and transcription regulation) and myopodin/Synpo2 (linking to vesicle formation and autophagy machinery). Mutations in the genes encoding various Z-disc proteins, in particular FLNc and its interaction partners, cause severe muscular diseases. We generated a knock-in mouse model mimicking the most prevalent human disease-causing FLNC mutation (p.W2710X) and found amplified lesion pathology as the major pathomechanism. In addition, we were also very successful in generating and analyzing mouse lines expressing BAG3-EGFP or the human pathogenic mutant BAG3P209L-EGFP in a muscle-specific and/or cardiac-specific manner. Cardiomyocytes expressing the pathogenic BAG3 mutant protein exhibit a phenotype mimicking the human pathology. These combined findings impressively illustrate the enormous importance of Z-disc protein-associated homeostasis mechanisms for muscle function and explain the dramatic impact of mutations affecting components of this machinery in protein aggregate myopathies.
Publications
- Distal myopathy with upper limb predominance caused by filamin C haploinsufficiency. Neurology 2011; 77:2105-2114
Guergueltcheva V, Peeters K, Baets J, Ceuterick-de Groote C, Martin JJ, Suls A, De Vriendt E, Mihaylova V, Chamova T, Almeida-Souza L, Ydens E, Tzekov C, Hadjidekov G, Gospodinova M, Storm K, Reyniers E, Bichev S, van der Ven PFM, Fürst DO, Mitev V, Lochmüller H, Timmerman V, Tournev I, De Jonghe P, Jordanova A
(See online at https://doi.org/10.1212/wnl.0b013e31823dc51e) - Mutations in the N-terminal actin-binding domain of filamin C cause a distal myopathy. Am J Hum Genet 2011; 88:729-740
Duff RM, Tay V, Hackman P, Ravenscroft G, McLean C, Kennedy P, Steinbach A, Schöffler W, van der Ven PFM, Fürst DO, Song J, Djinović-Carugo K, Penttilä S, Raheem O, Reardon K, Malandrini A, Gambelli S, Villanova M, Nowak KJ, Williams DR, Landers JE, Brown RH Jr, Udd B, Laing NG
(See online at https://doi.org/10.1016/j.ajhg.2011.04.021) - Pathophysiology of protein aggregation and extended phenotyping in filaminopathy. Brain 2012; 135:2642-2660
Kley RA, Serdaroglu-Oflazer P, Leber Y, Odgerel Z, van der Ven PFM, Olivé M, Ferrer I, Onipe A, Mihaylov M, Bilbao JM, Lee HS, Höhfeld J, Djinović-Carugo K, Kong K, Tegenthoff M, Peters SA, Stenzel W, Vorgerd M, Goldfarb LG, Fürst DO
(See online at https://doi.org/10.1093/brain/aws200) - A combined laser microdissection and mass spectrometry approach reveals new disease relevant proteins accumulating in aggregates of filaminopathy patients. Mol Cell Proteomics 2013; 12:215-227
Kley RA, Maerkens A, Leber Y, Theis V, Schreiner A, van der Ven PFM, Uszkoreit J, Stephan C, Eulitz S, Euler N, Kirschner J, Müller K, Meyer HE, Tegenthoff M, Fürst DO, Vorgerd M, Müller T, Marcus K
(See online at https://doi.org/10.1074/mcp.M112.023176) - Cellular mechanotransduction relies on tension-induced and chaperone-assisted autophagy. Curr Biol 2013; 23:430-435
Ulbricht A, Eppler FJ, Tapia VE, van der Ven PF, Hampe N, Hersch N, Vakeel P, Stadel D, Haas A, Saftig P, Behrends C, Fürst DO, Volkmer R, Hoffmann B, Kolanus W, Höhfeld J
(See online at https://doi.org/10.1016/j.cub.2013.01.064) - Differential proteomic analysis of abnormal intramyoplasmic aggregates in desminopathy. J Proteomics 2013; 90:14-27
Maerkens A, Kley RA, Olivé M, Theis V, van der Ven PFM, Reimann J, Milting H, Schreiner A, Uszkoreit J, Eisenacher M, Barkovits K, Güttsches AK, Tonillo J, Kuhlmann K, Meyer HE, Schröder R, Tegenthoff M, Fürst DO, Müller T, Goldfarb LG, Vorgerd M, Marcus K
(See online at https://doi.org/10.1016/j.jprot.2013.04.026) - Identification of Xin-repeat proteins as novel ligands of the SH3 domains of nebulin and nebulette and analysis of their interaction during myofibril formation and remodeling. Mol Biol Cell 2013; 24:3215-3226
Eulitz S, Sauer F, Pelissier M-C, Boisguerin P, Molt S, Schuld J, Orfanos Z, Kley RA, Volkmer R, Wilmanns M, Kirfel G, van der Ven PFM, Fürst DO
(See online at https://doi.org/10.1091/mbc.E13-04-0202) - Myopodin is an F-actin bundling protein with multiple independent actin-binding regions. J Muscle Res Cell Motil 2013; 34:61-69
Linnemann A, Vakeel P, Bezerra E, Orfanos Z, Djinović-Carugo K, van der Ven PFM, Kirfel G, Fürst DO
(See online at https://doi.org/10.1007/s10974-012-9334-5) - Aciculin interacts with filamin C and Xin and is essential for myofibril assembly and maintenance. J Cell Sci 2014; 127:3578-3592
Molt S, Bührdel JB, Yakovlev S, Schein P, Orfanos Z, Kirfel G, Winter L, Wiche G, van der Ven PFM, Rottbauer W, Just S, Belkin AM, Fürst DO
(See online at https://doi.org/10.1242/jcs.152157) - HSP70 Binding Protein HSPBP1 Regulates Chaperone Expression at a Posttranslational Level and is Essential for Spermatogenesis. Mol Biol Cell 2014; 25:2260-2271
Rogon C, Ulbricht A, Hesse M, Alberti S, Vijayaraj P, Best D, Adams IR, Magin TM, Fleischmann BK, Höhfeld J
(See online at https://doi.org/10.1091/mbc.E14-02-0742) - Myofibrillar instability exacerbated by acute exercise in filaminopathy. Hum Mol Genet 2015; 24: 7207-7220
Chevessier F, Schuld J, Orfanos Z, Plank AC, Wolf L, Maerkens A, Unger A, Schlötzer-Schrehardt U, Kley RA, Von Hörsten S, Marcus K, Linke WA, Vorgerd M, van der Ven PFM, Fürst DO, Schröder R
(See online at https://doi.org/10.1093/hmg/ddv421) - Breaking sarcomeres by in vitro exercise. Sci Rep 2016; 6:19614
Orfanos Z, Gödderz MP, Soroka E, Gödderz T, Rumyantseva A, van der Ven PFM, Hawke TJ, Fürst DO
(See online at https://doi.org/10.1038/srep19614) - Filamin C is a highly dynamic protein associated with fast repair of myofibrillar microdamage. Hum Mol Genet 2016; 25:2776-2788
Leber Y, Ruparelia AA, Kirfel G, van der Ven PFM, Hoffmann B, Merkel R, Bryson-Richardson RJ, Fürst DO
(See online at https://doi.org/10.1093/hmg/ddw135) - Sarcomeric lesions and remodeling proximal to intercalated discs in overloadinduced cardiac hypertrophy. Exp Cell Res 2016; 348:95-105
Kebir S, Orfanos Z, Schuld J, Linhart M, Lamberz C, van der Ven PFM, Schrickel J, Kirfel G, Fürst DO, Meyer R
(See online at https://doi.org/10.1016/j.yexcr.2016.09.008) - Myofibrillar Z-discs are a protein phosphorylation hot spot with PKC modulating protein dynamics. Mol Cell Proteom 2017; 16:346-367
Reimann L, Wiese H, Leber Y, Schwäble AN, Rohland A, Knapp B, Peikert CD, Drepper F, van der Ven PFM, Radziwill G, Fürst DO, Warscheid B
(See online at https://doi.org/10.1074/mcp.M116.065425) - The cochaperone BAG3 coordinates protein synthesis and autophagy under mechanical strain through spatial regulation of mTORC1. BBA – Mol Cell Res 2017; 1864:62-75
Kathage B, Gehlert S, Ulbricht A, Lüdecke L, Tapia VE, Orfanos Z, Wenzel D, Bloch W, Volkmer R, Fleischmann BK, Fürst DO, Höhfeld J
(See online at https://doi.org/10.1016/j.bbamcr.2016.10.007) - The novel cardiac z-disc protein CEFIP regulates cardiomyocyte hypertrophy by modulating calcineurin signaling. J Biol Chem 2017; 292: 15180-15191
Dierck, F, Kuhn C, Rohr C, Hille S, Braune J, Sossalla S, Molt S, van der Ven PFM, Fürst DO, Frey N
(See online at https://doi.org/10.1074/jbc.M117.786764)
