Abstract
Syndecan-4 is a cell membrane proteoglycan composed of a transmembrane core protein and substituted glycosaminoglycan (GAG) and N-linked glycosylated (N-glycosylated) chains. The core protein has three domains: extracellular, transmembrane and cytoplasmic domains. The GAG and N-glycosylated chains and the cytoplasmic domain of syndecan-4, especially the amino acids: Ser178 and Tyr187 are critical in regulation of turkey satellite cell growth and development. How these processes are regulated is still unknown. The objective of the current study was to determine whether the syndecan-4 GAG and N-glycosylated chains and the cytoplasmic domain functions through modulating focal adhesion formation and apoptosis. Twelve mutant clones were generated: a truncated syndecan-4 without the cytoplasmic domain with or without GAG and N-glycosylated chains, and Ser178 and Tyr187 mutants with or without GAG and N-glycosylated chains. The wild type syndecan-4 and all of the syndecan-4 mutants were transfected into turkey myogenic satellite cells after which cell apoptosis and focal adhesion formation were measured. Syndecan-4 increased cell membrane localization of β1 integrin and the activity of focal adhesion kinase (FAK) whereas the cytoplasmic domain mutation decreased the phosphorylation of FAK. However, syndecan-4 and syndecan-4 mutants did not influence cell apoptosis. They also had no effect on vinculin or paxillin-containing focal adhesion formation. These results suggested that the syndecan-4 cytoplasmic domain plays an important role in regulating FAK activity and β1 integrin cell membrane localization but not cell apoptosis and vinculin or paxillin-containing focal adhesion formation.
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References
Rapraeger AC (2001) Molecular interactions of syndecans during development. Semin Cell Dev Biol 12:107–116. doi:10.1006/scdb.2000.0239
Bernfield M, Gotte M, Park PW, Reizes O, Fitzgerald ML, Lincecum J, Zako M (1999) Functions of cell surface heparan sulfate proteoglycans. Annu Rev Biochem 68:729–777. doi:10.1146/annurev.biochem.68.1.729
Carey DJ (1997) Syndecans: multifunctional cell-surface co-receptors. Biochem J 327:1–16
Elenius K, Jalkanen M (1994) Function of the syndecans-a family of cell surface proteoglycans. J Cell Sci 107:2975–2982
Cornelison DD, Filla MS, Stanley HM, Rapraeger AC, Olwin BB (2001) Syndecan-3 and syndecan-4 specifically mark skeletal muscle satellite cells and are implicated in satellite cell maintenance and muscle regeneration. Dev Biol 239:79–94. doi:10.1006/dbio.2001.0416
Liu C, McFarland DC, Nestor KE, Velleman SG (2006) Differential expression of membrane-associated heparan sulfate proteoglycans in the skeletal muscle of turkeys with different growth rates. Poult Sci 85:422–428
Tanaka KK, Hall JK, Troy AA, Cornelison DD, Majka SM, Olwin BB (2009) Syndecan-4-expressing muscle progenitor cells in the SP engraft as satellite cells during muscle regeneration. Cell Stem Cell 4:217–225. doi:10.1016/j.stem.2009.01.016
Sporer KR, Chiang W, Tempelman RJ, Ernst CW, Reed KM, Velleman SG, Strasburg GM (2011) Characterization of a 6 K oligonucleotide turkey skeletal muscle microarray. Anim Genet 42:75–82. doi:10.1111/j.1365-2052.2010.02085.x
Echtermeyer F, Baciu PC, Saoncella S, Ge Y, Goetinck PF (1999) Syndecan-4 core protein is sufficient for the assembly of focal adhesions and actin stress fibers. J Cell Sci 112:3433–3441
Echtermeyer F, Streit M, Wilcox-Adelman S, Saoncella S, Denhez F, Detmar M, Goetinck P (2001) Delayed wound repair and impaired angiogenesis in mice lacking syndecan-4. J Clin Invest 107:R9–R14. doi:10.1172/JCI10559
Jeong J, Han I, Lim Y, Kim J, Park I, Woods A, Couchman JR, Oh ES (2001) Rat embryo fibroblasts require both the cell-binding and the heparin-binding domains of fibronectin for survival. Biochem J 356:531–537
Burridge K, Chrzanowska-Wodnicka M (1996) Focal adhesions, contractility, and signaling. Annu Rev Cell Dev Biol 12:463–518. doi:10.1146/annurev.cellbio.12.1.463
Clark EA, Brugge JS (1995) Integrins and signal transduction pathways: the road taken. Science 268:233–239. doi:10.1126/science.7716514
Hynes RO (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69:11–25. doi:10.1016/0092-8674(92)90115-S
Woods A, Couchman JR, Johansson S, Hook M (1986) Adhesion and cytoskeletal organisation of fibroblasts in response to fibronectin fragments. EMBO J 5:665–670
Saoncella S, Echtermeyer F, Denhez F, Nowlen JK, Mosher DF, Robinson SD, Hynes RO, Goetinck PF (1999) Syndecan-4 signals cooperatively with integrins in a Rho-dependent manner in the assembly of focal adhesions and actin stress fibers. Proc Natl Acad Sci USA 96:2805–2810. doi:10.1073/pnas.96.6.2805
Lyon M, Rushton G, Askari JA, Humphries MJ, Gallagher JT (2000) Elucidation of the structural features of heparan sulfate important for interaction with the Hep-2 domain of fibronectin. J Biol Chem 275:4599–4606. doi:10.1074/jbc.275.7.4599
Woods A, Longley RL, Tumova S, Couchman JR (2000) Syndecan-4 binding to the high affinity heparin-binding domain of fibronectin drives focal adhesion formation in fibroblasts. Arch Biochem Biophys 374:66–72. doi:10.1006/abbi.1999.1607
Longley RL, Woods A, Fleetwood A, Cowling GJ, Gallagher JT, Couchman JR (1999) Control of morphology, cytoskeleton and migration by syndecan-4. J Cell Sci 112:3421–3431
Oh ES, Woods A, Lim ST, Theibert AW, Couchman JR (1998) Syndecan-4 proteoglycan cytoplasmic domain and phosphatidylinositol 4,5-bisphosphate coordinately regulate protein kinase C activity. J Biol Chem 273:10624–10629. doi:10.1074/jbc.273.17.10624
Horowitz A, Murakami M, Gao Y, Simons M (1999) Phosphatidylinositol-4,5-bisphosphate mediates the interaction of syndecan-4 with protein kinase C. Biochemistry 38:15871–15877. doi:10.1021/bi991363i
Couchman JR, Vogt S, Lim ST, Lim Y, Oh ES, Prestwich GD, Theibert A, Lee W, Woods A (2002) Regulation of inositol phospholipid binding and signaling through syndecan-4. J Biol Chem 277:49296–49303. doi:10.1074/jbc.M209679200
Oh ES, Woods A, Couchman JR (1997) Syndecan-4 proteoglycan regulates the distribution and activity of protein kinase C. J Biol Chem 272:8133–8136. doi:10.1074/jbc.272.13.8133
Horowitz A, Simons M (1998) Phosphorylation of the cytoplasmic tail of syndecan-4 regulates activation of protein kinase C alpha. J Biol Chem 273:25548–25551. doi:10.1074/jbc.273.40.25548
Sun XG, Rotenberg SA (1999) Overexpression of protein kinase C alpha in MCF-10A human breast cells engenders dramatic alterations in morphology, proliferation, and motility. Cell Growth Differ 10:343–352
Murakami M, Horowitz A, Tang S, Ware JA, Simons M (2002) Protein kinase C (PKC) delta regulates PKC alpha activity in a syndecan-4-dependent manner. J Biol Chem 277:20367–20371. doi:10.1074/jbc.M202501200
Song Y, McFarland DC, Velleman SG (2012) Fibroblast growth factor 2 and protein kinase C alpha are involved in syndecan-4 cytoplasmic domain modulation of turkey myogenic satellite cell proliferation. Comp Biochem Physiol A 161:44–52. doi:10.1016/j.cbpa.2011.09.001
Song Y, McFarland DC, Velleman SG (2011) Role of syndecan-4 side chains in turkey satellite cell growth and development. Dev Growth Differ 53:97–109. doi:10.1111/j.1440-169X.2010.01230.x
Song Y, McFarland DC, Velleman SG (2012) Critical amino acids in syndecan-4 cytoplasmic domain modulation of turkey satellite cell growth and development. Comp Biochem Physiol A 161:271–278. doi:10.1016/j.cbpa.2011.11.007
Velleman SG, Liu X, Nestor KE, McFarland DC (2000) Heterogeneity in growth and differentiation characteristics in male and female satellite cells isolated from turkey lines with different growth rates. Comp Biochem Physiol A 125:503–509. doi:10.1016/S1095-6433(00)00178-1
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature (Lond) 237:680–685. doi:10.1038/227680a0
LeBaron RG, Esko JD, Woods A, Johansson S, Hook M (1988) Adhesion of glycosaminoglycan-deficient Chinese hamster ovary cell mutants to fibronectin substrata. J Cell Biol 106:945–952
Lim ST, Longley RL, Couchman JR, Woods A (2003) Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. J Biol Chem 278:13795–13802. doi:10.1074/jbc.M208300200
Keum E, Kim Y, Kim J, Kwon S, Lim Y, Han I, Oh ES (2004) Syndecan-4 regulates localization, activity and stability of protein kinase C-alpha. Biochem J 378:1007–1014. doi:10.1042/BJ20031734
Guan JL, Trevithick JE, Hynes RO (1991) Fibronectin/integrin interaction induces tyrosine phosphorylation of a 120-kDa protein. Cell Regul 2:951–964
Kornberg L, Earp HS, Parsons JT, Schaller M, Juliano RL (1992) Cell adhesion or integrin clustering increases phosphorylation of a focal adhesion-associated tyrosine kinase. J Biol Chem 267:23439–23442
Horowitz A, Simons M (1998) Regulation of syndecan-4 phosphorylation in vivo. J Biol Chem 273:10914–10918. doi:10.1074/jbc.273.18.10914
Mould AP, Humphries MJ (2004) Regulation of integrin function through conformational complexity: Not simply a knee-jerk reaction? Curr Opin Cell Biol 16:544–551
Giancotti FG, Ruoslahti E (1999) Integrin signaling. Science 285:1028–1033. doi:10.1126/science.285.5430.1028
Wilcox-Adelman SA, Denhez F, Goetinck PF (2002) Syndecan-4 modulates focal adhesion kinase phosphorylation. J Biol Chem 277:32970–32977. doi:10.1074/jbc.M201283200
Acknowledgments
Salary and partial research support to S.G.V. were provided by state and federal funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University, the Midwest Poultry Consortium, and the National Institute of Food and Agriculture Multistate Project NC-1184 to S.G.V. and D.C.M., and this project was supported by National Research Initiative Competitive Grant No. 2009-35503-05176 to S.G.V. from the USDA National Institute of Food and Agriculture.
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Song, Y., McFarland, D.C. & Velleman, S.G. Syndecan-4 cytoplasmic domain regulation of turkey satellite cell focal adhesions and apoptosis. Mol Biol Rep 39, 8251–8264 (2012). https://doi.org/10.1007/s11033-012-1673-1
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DOI: https://doi.org/10.1007/s11033-012-1673-1