SSeCKS promoted lipopolysaccharide-sensitized astrocytes migration via increasing β-1,4-galactosyltransferase-I activity
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Astrocytes migration is essential in the formation of the glial scar during the injury response process of the central nervous system (CNS) especially during inflammation. Integrin β1 is part of the extracellular matrix receptors in the CNS and it has been reported that integrin β-deficient astrocytes randomly migrate into wounds. Previous studies have found that β-1,4 Galactosyltransferase-I (β-1,4-GalT-I) enhanced the β-1,4-galactosylation of integrin β1. Src-suppressed C kinase substrate (SSeCKS) is an inflammatory response protein which functionally interacts with β-1,4 Galactosyltransferase-I (β-1,4-GalT-I). In this study we aim to investigate the role of SSeCKS and β-1,4-GalT-I in the migration of astrocytes during lipopolysaccharide (LPS)-induced inflammation. Coimmunoprecipitation and immunofluorescence assays have demonstrated that SSeCKS and β-1,4-GalT-I were significantly enhanced in LPS-treated astrocytes and their interactions may occur in the Trans-Golgi Network. Lectin blot showed that the knockdown of β-1,4-GalT-I could inhibit the β-1,4-galactosylation of glycoproteins including integrin β1 with and without LPS, and that SSeCKS knockdown inhibits the β-1,4-galactosylation of glycoproteins including integrin β1 only in LPS-induced astrocytes. Additionally, wound healing assays indicated that β-1,4-GalT-I knockdown could inhibit astrocytes migration with and without LPS but SSeCKS inhibited cell migration only when LPS was present. Therefore our findings suggest that SSeCKS affects astrocytes migration by regulating the β-1,4-galactosylation of glycoproteins including integrin β1, via β-1,4-GalT-I expression in LPS-sensitized astrocytes.
KeywordsAstrocytes Migration SSeCKS β-1,4-Galactosyltransferase-I Lipopolysaccharide
This work was supported by the National Natural Science Foundation of China (Grant No. 31600402), China Postdoctoral Science Fund (Grant No. 2017M621895), Zhejiang Province Postdoctoral Research Fund (Grant No. ZX2016000849), Ningbo Postdoctoral Research Fund (Grant No. ZX2017000053), Natural Science Foundation of Ningbo (Grant No. 2017A610216, 2016A610205), Natural Science Foundation of Zhejiang Province (Grant No. Y16H070001), Research Project of Zhejiang Provincial Department of Education (Y201738586), Agricultural Project of Public Welfare Technology Research in Zhejiang Provincial Science and Technology Department (Grant No. ZX2014C32047).
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Conflict of interest
The authors declare no conflict of interest.
- 8.Vogelezang M, Forster UB, Han J, Ginsberg MH, ffrench-Constant C (2007) Neurite outgrowth on a fibronectin isoform expressed during peripheral nerve regeneration is mediated by the interaction of paxillin with alpha4beta1 integrins. BMC Neurosci 8:44. https://doi.org/10.1186/1471-2202-8-44. doi: 1471-2202-8-44CrossRefGoogle Scholar
- 11.Guo HB, Lee I, Kamar M, Akiyama SK, Pierce M (2002) Aberrant N-glycosylation of beta1 integrin causes reduced alpha5beta1 integrin clustering and stimulates cell migration. Cancer Res 62(23):6837–6845Google Scholar
- 14.Owatworakit A, Townsend B, Louveau T, Jenner H, Rejzek M, Hughes RK, Saalbach G, Qi X, Bakht S, Roy AD, Mugford ST, Goss RJ, Field RA, Osbourn A (2013) Glycosyltransferases from oat (Avena) implicated in the acylation of avenacins. J Biol Chem 288(6):3696–3704. https://doi.org/10.1074/jbc.M112.42615. 5M112.426155 [pii]CrossRefGoogle Scholar
- 15.Asano M, Furukawa K, Kido M, Matsumoto S, Umesaki Y, Kochibe N, Iwakura Y (1997) Growth retardation and early death of beta-1,4-galactosyltransferase knockout mice with augmented proliferation and abnormal differentiation of epithelial cells. EMBO J 16(8):1850–1857. https://doi.org/10.1093/emboj/16.8.1850 CrossRefGoogle Scholar
- 16.Chui D, Oh-Eda M, Liao YF, Panneerselvam K, Lal A, Marek KW, Freeze HH, Moremen KW, Fukuda MN, Marth JD (1997) Alpha-mannosidase-II deficiency results in dyserythropoiesis and unveils an alternate pathway in oligosaccharide biosynthesis. Cell 90(1):157–167. doi:S0092-8674(00)80322-0 [pii]CrossRefGoogle Scholar
- 17.Yoshida A, Kobayashi K, Manya H, Taniguchi K, Kano H, Mizuno M, Inazu T, Mitsuhashi H, Takahashi S, Takeuchi M, Herrmann R, Straub V, Talim B, Voit T, Topaloglu H, Toda T, Endo T (2001) Muscular dystrophy and neuronal migration disorder caused by mutations in a glycosyltransferase, POMGnT1. Dev Cell 1(5):717–724. doi:S1534-5807(01)00070-3 [pii]CrossRefGoogle Scholar
- 22.Wassler MJ, Foote CI, Gelman IH, Shur BD (2001) Functional interaction between the SSeCKS scaffolding protein and the cytoplasmic domain of beta1,4-galactosyltransferase. J Cell Sci 114(Pt 12):2291–2300Google Scholar
- 23.Shao B, Li C, Yang H, Shen A, Wu X, Yuan Q, Kang L, Liu Z, Zhang G, Lu X, Cheng C (2011) The relationship between Src-suppressed C kinase substrate and beta-1,4 galactosyltransferase-I in the process of lipopolysaccharide-induced TNF-alpha secretion in rat primary astrocytes. Cell Mol Neurobiol 31(7):1047–1056. https://doi.org/10.1007/s10571-011-9704-3 CrossRefGoogle Scholar
- 26.Barry ST, Critchley DR (1994) The RhoA-dependent assembly of focal adhesions in Swiss 3T3 cells is associated with increased tyrosine phosphorylation and the recruitment of both pp125FAK and protein kinase C-delta to focal adhesions. J Cell Sci 107(Pt 7):2033–2045Google Scholar
- 27.Burnworth B, Pippin J, Karna P, Akakura S, Krofft R, Zhang G, Hudkins K, Alpers CE, Smith K, Shankland SJ, Gelman IH, Nelson PJ (2012) SSeCKS sequesters cyclin D1 in glomerular parietal epithelial cells and influences proliferative injury in the glomerulus. Lab Invest 92(4):499–510. https://doi.org/10.1038/labinvest.2011.199. labinvest2011199 [pii]CrossRefGoogle Scholar
- 30.Adams JC, Watt FM (1993) Regulation of development and differentiation by the extracellular matrix. Development 117(4):1183–1198Google Scholar
- 33.Isaji T, Sato Y, Fukuda T, Gu J (2009) N-glycosylation of the I-like domain of beta1 integrin is essential for beta1 integrin expression and biological function: identification of the minimal N-glycosylation requirement for alpha5beta1. J Biol Chem 284(18):12207–12216. https://doi.org/10.1074/jbc.M807920200 M807920200 [pii]CrossRefGoogle Scholar