Volume 10, Issue 2 (Vol.10 No.2 Jul 2021)                   rbmb.net 2021, 10(2): 145-155 | Back to browse issues page

XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Fahad Alharbi B, Al-Fahad D, Richard Dash P. Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells. rbmb.net. 2021; 10 (2) :145-155
URL: http://rbmb.net/article-1-690-en.html
Department of Pathological Analysis, College of Science, University of Thi-Qar, Thi-Qar, 64001 Iraq.
Abstract:   (737 Views)
Background: Focal adhesion (FA) play a critical role in many biological processes which include cell survival and cell migration. They serve as a cellular anchor, allowing cells to stay attached to the extracellular matrix (ECM), and can also regulate cellular transduction. Previously, it has been suggested that vesicles such as endosomes could interact directly with FA or be implicated in their turnover. In this study, we investigated whether there is a relationship between FA and the early endocytic machinery in MDA-MB-231 cells.

Methods: In this study, cell culture, transfection, time lapse confocal microscopies, immunocytochemistry, western blotting, cell fractionation, and immunoprecipitation techniques were performed.

Results: Cells acutely treated with Dynasore, an inhibitor of dynamin, or with Pitstop 2, an inhibitor of clathryn-dependent endocytosis showed a reduction in the expression of early endosome biomarkers such as Rab5 and EEA1. Additionally, cells treated with these endocytic inhibitors exhibited an increased number and size of FA, as well as an increased FA turnover duration. This data was consistent with the reduction of the speed of cell migration. We demonstrated that Rab5- and EEA1-positive early endosomes were found to be colocalized with internalized FA.

Conclusions: The present study suggests that there is a link between FA and early endosome markers, which indicates that the early endosomes may be involved in FA dynamics.                                  
Full-Text [PDF 319 kb]   (276 Downloads)    
Type of Article: Original Article | Subject: Cell Biology
Received: 2021/04/21 | Accepted: 2021/04/28 | Published: 2021/08/26

References
1. Murphy JE, Padilla BE, Hasdemir B, Cottrell GS, Bunnett NW. Endosomes: a legitimate platform for the signaling train. Proc Natl Acad Sci U S A. 2009;106(42):17615-22. [DOI:10.1073/pnas.0906541106] [PMID] [PMCID]
2. Tomas A, Futter CE, Eden ER. EGF receptor trafficking: consequences for signaling and cancer. Trends Cell Biol. 2014;24(1):26-34. [DOI:10.1016/j.tcb.2013.11.002] [PMID] [PMCID]
3. Maxfield FR, McGraw TE. Endocytic recycling. Nature Reviews Molecular Cell Biology. 2004;5(2):121-132. [DOI:10.1038/nrm1315] [PMID]
4. Helenius A, Mellman I, Wall D, Hubbard A. Endosomes. Trends in Biochemical Sciences. 1983;8(7):245-50. [DOI:10.1016/0968-0004(83)90350-X]
5. Mayor S, Pagano RE. Pathways of clathrin-independent endocytosis. Nature Reviews Molecular Cell Biology. 2007;8(8):603-612. [DOI:10.1038/nrm2216] [PMID]
6. Huotari J, Helenius A. Endosome maturation. EMBO J. 2011;30(17):3481-3500. [DOI:10.1038/emboj.2011.286] [PMID] [PMCID]
7. Wozniak MA, Modzelewska K, Kwong L, Keely PJ. Focal adhesion regulation of cell behavior. Biochimica et Biophysica Acta. 2004;1692(2-3):103-19. [DOI:10.1016/j.bbamcr.2004.04.007] [PMID]
8. Burridge K, Fath K, Kelly T, Nuckolls G, Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487-525. [DOI:10.1146/annurev.cb.04.110188.002415] [PMID]
9. Giannone G, Mège R-M, Thoumine O. Multi-level molecular clutches in motile cell processes. Trends Cell Biol. 2009; 19(9):475-86. [DOI:10.1016/j.tcb.2009.07.001] [PMID]
10. Meighan CM, Schwarzbauer JE. Temporal and spatial regulation of integrins during development. Curr Opin Cell Biol. 2008;20(5):520-524. [DOI:10.1016/j.ceb.2008.05.010] [PMID] [PMCID]
11. Webb DJ, Parsons JT, Horwitz AF. Adhesion assembly, disassembly and turnover in migrating cells - over and over and over again. Nature Cell Biology. 2002;4(4):E97-100. [DOI:10.1038/ncb0402-e97] [PMID]
12. Alfahad D, Alharethi S, Alharbi B, Mawlood K, Dash P. PtdIns(4,5)P2 and PtdIns(3,4,5)P3 dynamics during focal adhesions assembly and disassembly in a cancer cell line. Turk J Biol. 2020; 44(6):381-392. [DOI:10.3906/biy-2004-108] [PMID] [PMCID]
13. Meister M, Zuk A, Tikkanen R. Role of dynamin and clathrin in the cellular trafficking of flotillins. FEBS J. 2014;281(13):2956-76. [DOI:10.1111/febs.12834] [PMID]
14. Lees JG, Gorgani NN, Ammit AJ, McCluskey A, Robinson PJ, O'Neill GM. Role of dynamin in elongated cell migration in a 3D matrix. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2015;1853(3):611-618. [DOI:10.1016/j.bbamcr.2014.12.008] [PMID]
15. Skalski M, Sharma N, Williams K, Kruspe A, Coppolino MG. SNARE-mediated membrane traffic is required for focal adhesion kinase signaling and Src-regulated focal adhesion turnover. Biochim Biophys Acta. 2011;1813(1):148-58. [DOI:10.1016/j.bbamcr.2010.09.008] [PMID]
16. Chao W-T, Kunz J. Focal adhesion disassembly requires clathrin-dependent endocytosis of integrins. FEBS Letters. 2009;583(8):1337-43. [DOI:10.1016/j.febslet.2009.03.037] [PMID] [PMCID]
17. Ezratty EJ, Bertaux C, Marcantonio EE, Gundersen GG. Clathrin mediates integrin endocytosis for focal adhesion disassembly in migrating cells. J Cell Biol. 2009;187(5):733-47. [DOI:10.1083/jcb.200904054] [PMID] [PMCID]
18. Sadowski L, Pilecka I, Miaczynska M. Signaling from endosomes: Location makes a difference. Exp Cell Res. 2009;315(9):1601-9. [DOI:10.1016/j.yexcr.2008.09.021] [PMID]
19. Johnson IRD, Parkinson-Lawrence EJ, Keegan H, Spillane CD, Barry-O'Crowley J, Watson WR, et al. Endosomal gene expression: a new indicator for prostate cancer patient prognosis?. 2015;6(35):37919-37929. [DOI:10.18632/oncotarget.6114] [PMID] [PMCID]
20. Ohashi E, Tanabe K, Henmi Y, Mesaki K, Kobayashi Y, Takei K. Receptor Sorting within Endosomal Trafficking Pathway Is Facilitated by Dynamic Actin Filaments. PLoS One. 2011;6(5):e19942. [DOI:10.1371/journal.pone.0019942] [PMID] [PMCID]
21. Mettlen M, Pucadyil T, Ramachandran R, Schmid SL. Dissecting dynamin's role in clathrin-mediated endocytosis. Biochem Soc Trans. 2009;37(5):1022-6. [DOI:10.1042/BST0371022] [PMID] [PMCID]
22. Edeling M, Smith C, Owen D. Life of a clathrin coat: Insights from clathrin and AP structures. Nat Rev Mol Cell Biol. 2006;7(1):32-44. [DOI:10.1038/nrm1786] [PMID]
23. Von Kleist L, Stahlschmidt W, Bulut H, Gromova K, Puchkov D, Robertson MJ, et al. Role of the clathrin terminal domain in regulating coated pit dynamics revealed by small molecule inhibition. Cell. 2011;146(3):471-84. [DOI:10.1016/j.cell.2011.06.025] [PMID]
24. Wang Y, Cao H, Chen J, McNiven MA. A direct interaction between the large GTPase dynamin-2 and FAK regulates focal adhesion dynamics in response to active Src. Mol Biol Cell. 2011;22(9):1529-1538. [DOI:10.1091/mbc.e10-09-0785] [PMID] [PMCID]
25. Brown MC, Turner CE. Paxillin: Adapting to Change. Physiol Rev. 2004;84(4):1315-39. [DOI:10.1152/physrev.00002.2004] [PMID]
26. Côté J-F, Turner CE, Tremblay ML. Intact LIM 3 and LIM 4 Domains of Paxillin Are Required for the Association to a Novel Polyproline Region (Pro 2) of Protein-Tyrosine Phosphatase-PEST. J Biol Chem. 1999;274(29):20550-60. [DOI:10.1074/jbc.274.29.20550] [PMID]
27. Horton ER, Byron A, Askari JA, Ng DHJ, Millon-Frémillon A, Robertson J, et al. Definition of a consensus integrin adhesome and its dynamics during adhesion complex assembly and disassembly. Nat Cell Biol. 2015;17(12):1577-1587. [DOI:10.1038/ncb3257] [PMID] [PMCID]
28. Márquez MG, Brandán YR, Guaytima E del V, Paván CH, Favale NO, Sterin-Speziale NB. Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway. Biochim Biophys Acta. 2014;1843(12):2991-3003. [DOI:10.1016/j.bbamcr.2014.09.014] [PMID]
29. Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C, Kirchhausen T. Dynasore, a Cell-Permeable Inhibitor of Dynamin. Dev Cell. 2006;10(6):839-50. [DOI:10.1016/j.devcel.2006.04.002] [PMID]
30. Cleghorn WM, Branch KM, Kook S, Arnette C, Bulus N, Zent R, el at. Arrestins regulate cell spreading and motility via focal adhesion dynamics. Mol Biol Cell. 2015;26(4):622-635. [DOI:10.1091/mbc.E14-02-0740] [PMID] [PMCID]
31. Yoshida A, Sakai N, Uekusa Y, Imaoka Y, Itagaki Y, Suzuki Y, el at. Changes of plasma membrane and protein assembly during clathrin-mediated endocytosis. PLoS Biol. 2018;16(5):e2004786. [DOI:10.1371/journal.pbio.2004786] [PMID] [PMCID]
32. Rink J, Ghigo E, Kalaidzidis Y, Zerial M. Rab Conversion as a Mechanism of Progression from Early to Late Endosomes. Cell. 2005;122(5):735-49. [DOI:10.1016/j.cell.2005.06.043] [PMID]
33. Schiefermeier N, Scheffler JM, de Araujo MEG, Stasyk T, Yordanov T, Ebner HL el at. The late endosomal p14-MP1 (LAMTOR2/3) complex regulates focal adhesion dynamics during cell migration. J Cell Biol. 2014;205(4):525-40. [DOI:10.1083/jcb.201310043] [PMID] [PMCID]
34. Sharifi MN, Mowers EE, Drake LE, Collier C, Chen H, Zamora M, el at. Autophagy Promotes Focal Adhesion Disassembly and Cell Motility of Metastatic Tumor Cells through the Direct Interaction of Paxillin with LC3. Cell Rep. 2016;15(8):1660-72. [DOI:10.1016/j.celrep.2016.04.065] [PMID] [PMCID]
35. Roberts M, Barry S, Woods A, Sluijs P van der, Norman J. PDGF-regulated rab4-dependent recycling of αvβ3 integrin from early endosomes is necessary for cell adhesion and spreading. Curr Biol. 2001;11(18):1392-402. [DOI:10.1016/S0960-9822(01)00442-0]
36. Caswell P, Norman J. Endocytic transport of integrins during cell migration and invasion. Trends Cell Biol. 2008;18(6):257-63. [DOI:10.1016/j.tcb.2008.03.004] [PMID]

Add your comments about this article : Your username or Email:
CAPTCHA

Send email to the article author


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2015 All Rights Reserved | Reports of Biochemistry and Molecular Biology

Designed & Developed by : Yektaweb