Identification of Nucleolin as a Lipid-Raft-Dependent Beta1-Integrin-Interacting Protein in A375 Cell Migration

다국어 초록 (Multilingual Abstract)

Lipid rafts are related to cell surface receptor function. Integrin is a major surface receptor protein in cell adhesion and migration on the extracellular matrix (ECM). Here, we showed that lipid rafts played a critical role in human melanoma A375 cell spreading and migration on fibronectin; an important component of the ECM that interacts with 1 integrin. We found that the disruption of lipid rafts did not markedly inhibit the expression and activation of 1 integrin. By coimmunoprecipitation and mass spectrometry, we investigated the influence of lipid rafts on the 1 integrin complex and identified nucleolin as a potential lipid-raft-dependent 1-integrin-interacting protein. Upon confirmation of the interaction between 1 integrin and nucleolin, further studies revealed that nucleolin colocalized with 1 integrin in lipid rafts and raft disruption interrupted their association. In addition, knockdown of nucle-olin markedly attenuated A375 cell spreading and migration on fibronectin. Taken together, we demonstrated that nucleolin is a critical lipid-raft-dependent 1-integrin-inte-racting protein in A375 cell spreading and migration on fibronectin.

참고문헌 (Reference)

1 Riopel, M.M, "β1 integrin-extracellular matrix interactions are essential for maintaining exocrine pancreas architecture and function" 93 : 31-40, 2013

2 Dumler, I, "Urokinase- induced mitogenesis is mediated by casein kinase 2 and nucleolin" 9 : 1468-1476, 1999

3 Leitinger, B, "The involvement of lipid rafts in the regulation of integrin function" 115 : 963-972, 2002

4 Said, E.A, "The anti-HIV cytokine midkine binds the cell surface-expressed nucleolin as a low affinity receptor" 277 : 37492-37502, 2002

5 Huang, Y, "The angiogenic function of nucleolin is mediated by vascular endothelial growth factor and nonmuscle myosin" 107 : 3564-3571, 2006

6 Zhang, Z, "The alpha 5 beta 1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression" 92 : 6161-6165, 1995

7 El Khoury, D, "Targeting surface nucleolin with a multivalent pseudopeptide delays development of spontaneous melanoma in RET transgenic mice" 10 : 325-, 2010

8 Hovanessian, A.G, "Surface expressed nucleolin is constantly induced in tumor cells to mediate calcium-dependent ligand internalization" 5 : e15787-, 2010

9 Farin, K, "Structure- function analysis of nucleolin and ErbB receptors interactions" 4 : e6128-, 2009

10 Golub, T, "Spatial and temporal control of signaling through lipid rafts" 14 : 542-550, 2004

11 Hehlgans, S, "Signalling via integrins: implications for cell survival and anticancer strategies" 1775 : 163-180, 2007

12 Ikonen, E, "Roles of lipid rafts in membrane transport" 13 : 470-477, 2001

13 Yanagisawa, M, "Roles of lipid rafts in integrin-dependent adhesion and gp130 signalling pathway in mouse embryonic neural precursor cells" 9 : 801-809, 2004

14 Hood, J.D, "Role of integrins in cell invasion and migration" 2 : 91-100, 2002

15 Simons, K., "Revitalizing membrane rafts: new tools and insights" 11 : 688-699, 2011

16 Baron, W, "Regulation of integrin growth factor interactions in oligodendrocytes by lipid raft microdomains" 13 : 151-155, 2003

17 Chen, H.M, "Overexpression of integrin-β1 in leiomyoma promotes cell spreading and proliferation" 98 : E837-E846, 2013

18 Chen, X, "Nucleolinmediated cellular trafficking of DNA nanoparticle is lipid raft and microtubule dependent and can be modulated by glucocorticoid" 19 : 93-102, 2011

19 Holly, S.P, "Multiple roles of integrins in cell motility" 261 : 69-74, 2000

20 Legate, K.R, "Mechanisms that regulate adaptor binding to beta-integrin cytoplasmic tails" 122 : 187-198, 2009

21 Ogier, C, "Matrix metalloproteinase-2 (MMP-2) regulates astrocyte motility in connection with the actin cytoskeleton and integrins" 54 : 272-284, 2006

22 Meyer, S, "Lubrol-RAFTs in melanoma cells: a molecular platform for tumor-promoting ephrin-B2-integrin-beta1 interaction" 127 : 1615-1621, 2007

23 Vassilieva, E.V, "Lipid rafts mediate internalization of beta1-integrin in migrating intestinal epithelial cells" 295 : G965-G976, 2008

24 Mañes, S, "Lipid rafts in lymphocyte activation and migration" 23 : 59-69, 2006

25 Simons, K, "Lipid rafts and signal transduction" 1 : 31-39, 2000

26 Rajendran, L, "Lipid rafts and membrane dynamics" 118 : 1099-1102, 2005

27 Wang, R., "Lipid raft regulates the initial spreading of melanoma A375 cells by modulating β1 integrin clustering" 45 : 1679-1689, 2013

28 Jae-Hong Kim, "Leukotriene Synthesis in Response to A23187 Is Inhibited by Methyl- β-Cyclodextrin in RBL-2H3 Cells" 한국분자세포생물학회 23 (23): 57-63, 2007

29 Mao, L, "Label-free proteomics reveals decreased expression of CD18 and AKNA in peripheral CD4+ T cells from patients with Vogt-Koyanagi-Harada syndrome" 6 : e14616-, 2011

30 Koutsioumpa, M, "Interplay between αvβ3 integrin and nucleolin regulates human endothelial and glioma cell migration" 288 : 343-354, 2013

31 Porter, J.C, "Integrins take partners: cross-talk between integrins and other membrane receptors" 8 : 390-396, 1998

32 Danen, E.H, "Integrins control motile strategy through a Rho-cofilin pathway" 169 : 515-526, 2005

33 Barczyk, M, "Integrins" 339 : 269-280, 2010

34 Mostafavi-Pour, Z, "Integrin-specific signaling pathways controlling focal adhesion formation and cell migration" 161 : 155-167, 2003

35 Galzio, R, "Glycosilated nucleolin as marker for human gliomas" 113 : 571-579, 2012

36 Li, C, "Gambogic acid inhibits tumor cell adhesion by suppressing integrin β1 and membrane lipid raftsassociated integrin signaling pathway" 82 : 1873-1883, 2011

37 Wang, Y, "Cell-surface nucleolin is involved in lipopolysaccharide internalization and signalling in alveolar macrophages" 35 : 677-685, 2011

38 Reyes-Reyes, E.M, "Cell-surface nucle-olin is a signal transducing P-selectin binding protein for human colon carcinoma cells" 314 : 2212-2223, 2008

39 Maheshwari, G, "Cell adhesion and motility depend on nanoscale RGD clustering" 113 : 1677-1686, 2000

40 Kang, W.J, "Bioimaging of geographically adjacent proteins in a single cell by quantum dotbased fluorescent resonance energy transfer" 3 : 1383-1388, 2009

41 Nisole, S, "Anchorage of HIV on permissive cells leads to coaggregation of viral particles with surface nucleolin at membrane raft microdomains" 276 : 155-173, 2002

42 Villalba, M, "A Novel Functional Interaction between Vav and PKCu Is Required for TCR-Induced T Cell Activation" 12 : 151-160, 2000