Abstract
The intermediate filament protein desmin generates an extra-sarcomeric network in myocytes. Mutations in the desmin gene cause myofibrillar myopathy characterized by desmin-positive aggregates and myofibrillar dissolution. Past analysis revealed that the non-α-helical amino-terminal “head” domain of desmin is a vital coordinator of protein assembly. We have now characterized assembly and network-forming properties of five recently discovered myopathy-causing mutations residing in this domain. In vitro analyses with recombinant proteins show that two mutant variants residing in a conserved nonapeptide motif “SSYRRTFGG”—Ser13Phe and Arg16Cys—interfere with assembly by forming filamentous aggregates. Consistent with in vitro data, both mutant proteins are unable to generate a bona fide filament system in cells lacking an intermediate filament cytoskeleton. In cells expressing vimentin or desmin, both mutants firstly fail to integrate into the endogenous filament network and secondly severely affect its cellular localization. The other three mutations—Ser2Iso, Ser46Phe, and Ser46Tyr—influence in vitro filament properties less severely, but in vivo, Ser46Phe and Ser46Tyr impair de novo filament formation. These effects of the “head” mutant proteins on endogenous intermediate filament system and their competition for binding to cellular anchoring structures might explain part of the molecular mechanism that causes disease.
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References
Capetanaki Y, Bloch RJ, Kouloumenta A, Mavroidis M, Psarras S (2007) Muscle intermediate filaments and their links to membranes and membranous organelles. Exp Cell Res 313:2063–2076
Bär H, Strelkov SV, Sjöberg G, Aebi U, Herrmann H (2004) The biology of desmin filaments: how do mutations affect their structure, assembly, and organisation? J Struct Biol 148:137–152
Herrmann H, Bär H, Kreplak L, Strelkov SV, Aebi U (2007) Intermediate filaments: from cell architecture to nanomechanics. Nat Rev Mol Cell Biol 8:562–573
Goldfarb LG, Vicart P, Goebel HH, Dalakas MC (2004) Desmin myopathy. Brain 127:723–734
Taylor MR, Slavov D, Ku L, Di Lenarda A, Sinagra G, Carniel E, Haubold K, Boucek MM, Ferguson D, Graw SL, Zhu X, Cavanaugh J, Sucharov CC, Long CS, Bristow MR, Lavori P, Mestroni L (2007) Prevalence of desmin mutations in dilated cardiomyopathy. Circulation 115:1244–1251
Herrmann H, Aebi U (1998) Structure, assembly, and dynamics of intermediate filaments. Subcell Biochem 31:319–362
Pruszczyk P, Kostera-Pruszczyk A, Shatunov A, Goudeau B, Draminska A, Takeda K, Sambuughin N, Vicart P, Strelkov SV, Goldfarb LG, Kaminska A (2007) Restrictive cardiomyopathy with atrioventricular conduction block resulting from a desmin mutation. Int J Cardiol 117:244–253
Arbustini E, Pasotti M, Pilotto A, Pellegrini C, Grasso M, Previtali S, Repetto A, Bellini O, Azan G, Scaffino M, Campana C, Piccolo G, Vigano M, Tavazzi L (2006) Desmin accumulation restrictive cardiomyopathy and atrioventricular block associated with desmin gene defects. Eur J Heart Fail 8:477–483
Bär H, Goudeau B, Wälde S, Casteras-Simon M, Mücke N, Shatunov A, Goldberg YP, Clarke C, Holton JL, Eymard B, Katus HA, Fardeau M, Goldfarb L, Vicart P, Herrmann H (2007) Conspicuous involvement of desmin tail mutations in diverse cardiac and skeletal myopathies. Hum Mutat 28:374–386
Selcen D, Ohno K, Engel AG (2004) Myofibrillar myopathy: clinical, morphological and genetic studies in 63 patients. Brain 127:439–451
Bergman JE, Veenstra-Knol HE, van Essen AJ, van Ravenswaaij CM, den Dunnen WF, van den Wijngaard A, van Tintelen JP (2007) Two related Dutch families with a clinically variable presentation of cardioskeletal myopathy caused by a novel S13F mutation in the desmin gene. Eur J Med Genet 50:355–366
Pica EC, Kathirvel P, Pramono ZA, Lai PS, Yee WC (2008) Characterization of a novel S13F desmin mutation associated with desmin myopathy and heart block in a Chinese family. Neuromuscul Disord 18:178–182
Olive M, Armstrong J, Miralles F, Pou A, Fardeau M, Gonzalez L, Martinez F, Fischer D, Martinez Matos JA, Shatunov A, Goldfarb L, Ferrer I (2007) Phenotypic patterns of desminopathy associated with three novel mutations in the desmin gene. Neuromuscul Disord 17:443–450
Bär H, Mücke N, Kostareva A, Sjöberg G, Aebi U, Herrmann H (2005) Severe muscle disease-causing desmin mutations interfere with in vitro filament assembly at distinct stages. Proc Natl Acad Sci U S A 102:15099–15104
Bär H, Kostareva A, Sjöberg G, Sejersen T, Katus HA, Herrmann H (2006) Forced expression of desmin and desmin mutants in cultured cells: impact of myopathic missense mutations in the central coiled-coil domain on network formation. Exp Cell Res 312:1554–1565
Herrmann H, Haner M, Brettel M, Müller SA, Goldie KN, Fedtke B, Lustig A, Franke WW, Aebi U (1996) Structure and assembly properties of the intermediate filament protein vimentin: the role of its head, rod and tail domains. J Mol Biol 264:933–953
Beuttenmuller M, Chen M, Janetzko A, Kuhn S, Traub P (1994) Structural elements of the amino-terminal head domain of vimentin essential for intermediate filament formation in vivo and in vitro. Exp Cell Res 213:128–142
Schaffeld M, Herrmann H, Schultess J, Markl J (2001) Vimentin and desmin of a cartilaginous fish, the shark Scyliorhinus stellaris: sequence, expression patterns and in vitro assembly. Eur J Cell Biol 80:692–702
Granger BL, Lazarides E (1980) Synemin: a new high molecular weight protein associated with desmin and vimentin filaments in muscle. Cell 22:727–738
Wickert U, Mücke N, Wedig T, Muller SA, Aebi U, Herrmann H (2005) Characterization of the in vitro co-assembly process of the intermediate filament proteins vimentin and desmin: mixed polymers at all stages of assembly. Eur J Cell Biol 84:379–391
Mücke N, Wedig T, Burer A, Marekov LN, Steinert PM, Langowski J, Aebi U, Herrmann H (2004) Molecular and biophysical characterization of assembly-starter units of human vimentin. J Mol Biol 340:97–114
Herrmann H, Kreplak L, Aebi U (2004) Isolation, characterization, and in vitro assembly of intermediate filaments. Meth Cell Biol 78:3–24
Bär H, Fischer D, Goudeau B, Kley RA, Clemen CS, Vicart P, Herrmann H, Vorgerd M, Schröder R (2005) Pathogenic effects of a novel heterozygous R350P desmin mutation on the assembly of desmin intermediate filaments in vivo and in vitro. Hum Mol Genet 14:1251–1260
Colucci-Guyon E, Portier MM, Dunia I, Paulin D, Pournin S, Babinet C (1994) Mice lacking vimentin develop and reproduce without an obvious phenotype. Cell 79:679–694
Claycomb WC, Lanson NA Jr, Stallworth BS, Egeland DB, Delcarpio JB, Bahinski A, Izzo NJ Jr (1998) HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. Proc Natl Acad Sci U S A 95:2979–2984
Parry DA, Steinert PM (1995) Intermediate filament structure. Springer, New York
Bär H, Mücke N, Ringler P, Müller SA, Kreplak L, Katus HA, Aebi U, Herrmann H (2006) Impact of disease mutations on the desmin filament assembly process. J Mol Biol 360:1031–1042
Herrmann H, Hofmann I, Franke WW (1992) Identification of a nonapeptide motif in the vimentin head domain involved in intermediate filament assembly. J Mol Biol 223:637–650
Goldfarb L, Olive M, Vicart P, Goebel HH (2008) Intermediate filament diseases: desminopathy. Adv Exp Med Biol 642:131–164
Kaufmann E, Weber K, Geisler N (1985) Intermediate filament forming ability of desmin derivatives lacking either the amino-terminal 67 or the carboxy-terminal 27 residues. J Mol Biol 185:733–742
Matsuzawa K, Kosako H, Azuma I, Inagaki N, Inagaki M (1998) Possible regulation of intermediate filament proteins by Rho-binding kinases. Subcell Biochem 31:423–435
Parry DA (2005) Microdissection of the sequence and structure of intermediate filament chains. Adv Protein Chem 70:113–142
Kreplak L, Bär H (2009) Severe myopathy mutations modify the nanomechanics of desmin intermediate filaments. J Mol Biol 385:1043–1051
Clemen CS, Fischer D, Reimann J, Eichinger L, Müller CR, Müller HD, Goebel HH, Schröder R (2009) How much mutant protein is needed to cause a protein aggregate myopathy in vivo? Lessons from an exceptional desminopathy. Hum Mutat 30:E490–E499
Conover GM, Henderson SN, Gregorio CC (2009) A myopathy-linked desmin mutation perturbs striated muscle actin filament architecture. Mol Biol Cell 20:834–845
Acknowledgments
Harald Bär and Harald Herrmann acknowledge grants from the German Research Foundation (DFG; BA 2186/3-1 to H.B. and H.H.). Full-length cDNA of human LMNB1 cloned into the eukaryotic expression vector pEYFP-C1 was generously provided by Stephanie Geiger, DKFZ, Germany. The HL-1 cells were kindly provided by William C. Claycomb, Louisiana State University Medical Center, New Orleans, USA. We wish to thank Gloria Conover from Texas A&M University for critical reading of the manuscript.
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The authors declare that they have no conflict of interests.
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Sharma, S., Mücke, N., Katus, H.A. et al. Disease mutations in the “head” domain of the extra-sarcomeric protein desmin distinctly alter its assembly and network-forming properties. J Mol Med 87, 1207–1219 (2009). https://doi.org/10.1007/s00109-009-0521-9
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DOI: https://doi.org/10.1007/s00109-009-0521-9