Abstract
The nebulin family of actin-binding proteins plays an essential role in cytoskeletal dynamics and actin filament stability. All of the family members are modular proteins with their key defining structural feature being the presence of the 35-residue nebulin modules. The family members now include nebulin, nebulette, N-RAP, LASP-1, and LIM-nebulette. Nebulin and nebulette are associated with the thin filament/Z-line junction of striated muscle. LASP-1 and LIM-nebulette are found within focal adhesions, and N-RAP is associated with muscle cellular junctions. Although much investigation has focused on the role of the interactions between nebulin modules and actin, each of these proteins contains other domains that are essential for their cellular targeting and functions. The serine-rich linker region of nebulette has previously been shown to serve just such a purpose by targeting the association of the nebulin modules to the cardiac Z-line in cultured cardiomyocytes. In this report, we analyze the targeting functions of the homologous regions of LASP-1 and LIM-nebulette in their incorporation into focal adhesions. We have found that the linker region of LASP-1 is indeed important for its cellular localization and that the shortened linker region of LIM-nebulette drives the association of nebulin modules to focal adhesions.
Similar content being viewed by others
References
Arimura T, Nakamura T, Hiroi S, Satoh M, Takahashi M, Ohbuchi N, Ueda K, Nouchi T, Yamaguchi N, Akai J, Matsumori A, Sasayama S, Kimura A (2000) Characterization of the human nebulette gene: a polymorphism in an actin-binding motif is associated with nonfamilial idiopathic dilated cardiomyopathy. Hum Genet 107:440–451
Chew CS, Parente JA, Zhou C Jr, Baranco E, Chen X (1998) Lasp-1 is a regulated phosphoprotein within the cAMP signaling pathway in the gastric parietal cell. Am J Physiol 275:C56–C67
Chew CS, Parente JA, Chen X Jr, Chaponnier C, Cameron RS (2000) The LIM and SH3 domain-containing protein, Lasp-1, may link the cAMP signaling pathway with dynamic membrane restructuring activities in ion transporting epithelia. J Cell Sci 113:2035–2045
Chew CS, Chen X, Parente JA, Tarrer S Jr, Okamoto C, Qin HY (2002) Lasp-1 binds to non-muscle F-actin in vitro and is localized within multiple sites of dynamic actin assembly in vivo. J Cell Sci 115:4787–4799
Ehler E, Horowits R, Zuppinger C, Price RL, Perriard E, Leu M, Caroni P, Sussman M, Eppenberger HM, Perriard JC (2001) Alterations at the intercalated disk associated with the absence of muscle LIM protein. J Cell Biol 153:763–772
Grunewald TG, Kammerer U, Schulze E, Schindler D, Honig A, Zimmer M, Butt E (2006) Silencing of LASP-1 influences zyxin localization, inhibits proliferation and reduces migration in breast cancer cells. Exp Cell Res (in press)
He TC, Zhou S, Costa LT da, Yu J, Kinzler KW, Vogelstein B (1998) A simplified system for generating recombinant adenoviruses. Proc Nat Acad Sci USA 95:2509–2514
Jin JP, Wang K (1991) Cloning, expression, and protein interaction of human nebulin fragments composed of varying numbers of sequence modules. J Biol Chem 266:21215–21223
Katoh M (2003) Identification and characterization of LASP2 gene in silico. Int J Mol Med 12:405–410
Li B, Zhuang L, Trueb B (2004) Zyxin interacts with the SH3 domains of the cytoskeletal proteins LIM-nebulette and Lasp-1. J Biol Chem 279:20401–20410
Lin YH, Park ZY, Lin D, Brahmbhatt AA, Rio MC, Yates JR 3rd, Klemke RL (2004) Regulation of cell migration and survival by focal adhesion targeting of Lasp-1. J Cell Biol 165:421–432
Luo G, Zhang JQ, Nguyen TP, Herrera AH, Paterson B, Horowits R (1997) Complete cDNA sequence and tissue localization of N-RAP, a novel nebulin-related protein of striated muscle. Cell Motil Cytoskeleton 38:75–90
Luo G, Herrera AH, Horowits R (1999) Molecular interactions of N-RAP, a nebulin-related protein of striated muscle myotendon junctions and intercalated disks. Biochemistry 38:6135–6143
Moncman CL, Rindt H, Robbins J, Winkelmann DA (1993) Segregated assembly of muscle myosin expressed in nonmuscle cells. Mol Biol Cell 4:1051–1067
Moncman CL, Wang K (1995) Nebulette: a 107 kD nebulin-like protein in cardiac muscle. Cell Motil Cytoskeleton 32:205–225
Moncman CL, Wang K (1999) Functional dissection of nebulette demonstrates actin binding of nebulin-like repeats and Z-line targeting of SH3 and linker domains. Cell Motil Cytoskeleton 44:1–22
Moncman CL, Wang K (2000) Architecture of the thin filament Z-line junction: lessons from nebulette and nebulin homologies. J Muscle Res Cell Motil 21:153–169
Moncman CL, Wang K (2002) Targeted disruption of nebulette protein expression alters cardiac myofibril assembly and function. Exp Cell Res 273:204–218
Ojima K, Lin ZX, Bang M, Holtzer S, Matsuda R, Labeit S, Sweeney HL, Holtzer H (2000) Distinct families of Z-line targeting modules in the COOH-terminal region of nebulin. J Cell Biol 150:553–566
Pelin K, Hilpela P, Donner K, Sewry C, Akkari PA, Wilton SD, Wattanasirichaigoon D, Bang ML, Centner T, Hanefeld F, Odent S, Fardeau M, Urtizberea JA, Muntoni F, Dubowitz V, Beggs AH, Laing NG, Labeit S, Chapelle A de la, Wallgren-Pettersson C (1999) Mutations in the nebulin gene associated with autosomal recessive nemaline myopathy. Proc Nat Acad Sci USA 96:2305–2310
Pfuhl M, Winder SJ, Pastore A (1994) Nebulin, a helical actin binding protein. EMBO J 13:1782–1789
Pfuhl M, Winder SJ, Castiglione Morelli MA, Labeit S, Pastore A (1996) Correlation between conformational and binding properties of nebulin repeats. J Mol Biol 257:367–384
Rhee D, Sanger JM, Sanger JW (1994) The premyofibril: evidence for its role in myofibrillogenesis. Cell Motil Cytoskeleton 28:1–24
Schreiber V, Masson R, Linares JL, Mattei MG, Tomasetto C, Rio MC (1998a) Chromosomal assignment and expression pattern of the murine Lasp-1 gene. Gene 207:171–175
Schreiber V, Moog-Lutz C, Regnier CH, Chenard MP, Boeuf H, Vonesch JL, Tomasetto C, Rio MC (1998b) Lasp-1, a novel type of actin-binding protein accumulating in cell membrane extensions. Mol Med 4:675–687
Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653–1663
Somerville LL, Wang K (1987) In vivo phosphorylation of titin and nebulin in frog skeletal muscle. Biochem Biophys Res Commun 147:986–992
Somerville LL, Wang K (1988) Sarcomere matrix of striated muscle: in vivo phosphorylation of titin and nebulin in mouse diaphragm muscle. Arch Biochem Biophys 262:118–129
Spence HJ, McGarry L, Chew CS, Carragher NO, Scott-Carragher LA, Yuan Z, Croft DR, Olson MF, Frame M, Ozanne BW (2006) AP-1 differentially expressed proteins Krp1 and fibronectin cooperatively enhance Rho-ROCK-independent mesenchymal invasion by altering the function, localization, and activity of nondifferentially expressed proteins. Mol Cell Biol 26:1480–1495
Terasaki AG, Suzuki H, Nishioka T, Matsuzawa E, Katsuki M, Nakagawa H, Miyamoto S, Ohashi K (2004) A novel LIM and SH3 protein (lasp-2) highly expressing in chicken brain. Biochem Biophys Res Commun 313:48–54
Tomasetto C, Moog-Lutz C, Regnier CH, Schreiber V, Basset P, Rio MC (1995) Lasp-1 (MLN 50) defines a new LIM protein subfamily characterized by the association of LIM and SH3 domains. FEBS Lett 373:245–249
van der Ven PF, Jap PH, Laak HJ ter, Nonaka I, Barth PG, Sengers RC, Stadhouders AM, Ramaekers FC (1995) Immunophenotyping of congenital myopathies: disorganization of sarcomeric, cytoskeletal and extracellular matrix proteins. J Neurol Sci 129:199–213
Wang K, Williamson CL (1980) Identification of an N2 line protein of striated muscle. Proc Nat Acad Sci USA 77:3254–3258
Wang K, Knipfer M, Huang QQ, Heerden A van, Hsu LC, Gutierrez G, Quian XL, Stedman H (1996) Human skeletal muscle nebulin sequence encodes a blueprint for thin filament architecture. Sequence motifs and affinity profiles of tandem repeats and terminal SH3. J Biol Chem 271:4304–4314
Acknowledgements
The authors are indebted to Mr. Michael Esham for technical assistance, to Drs. Rick McCann and Kathleen Doane for critical reading of this manuscript, and to Dr. Kuan Wang for continuing support and encouragement of this work.
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by grants from the National Institutes of Health-HLB and the National Council of the American Heart Association to C.L.M.
Rights and permissions
About this article
Cite this article
Panaviene, Z., Moncman, C.L. Linker region of nebulin family members plays an important role in targeting these molecules to cellular structures. Cell Tissue Res 327, 353–369 (2007). https://doi.org/10.1007/s00441-006-0305-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-006-0305-2