Cell Spreading Area Regulates Clathrin-coated Pit Dynamics on Micropatterned Substrate

Overview

Journal Integr Biol (Camb)

Publisher Oxford University Press

Specialty Biology

Date 2015 Jul 25

PMID 26205141

Citations 21

Authors

Xinyu Tan

Johanna Heureaux

Allen P Liu

Affiliations

Soon will be listed here.

Abstract

Clathrin-mediated endocytosis (CME) is the most characterized pathway for the endocytic entry of proteins and lipids at the plasma membrane of eukaryotic cells. Numerous studies have probed the roles of different endocytic accessory proteins in regulating the dynamics of clathrin-coated pit (CCP) assembly. However, it is not completely clear how physical cues regulate CCP dynamics. Here we employ microcontact printing to control cell shape and examine CCP dynamics as a function of cell spreading area for three differently sized cells. Cells with a large spreading area had more short-lived CCPs but a higher CCP initiation rate. Interestingly, we found that fluorescence intensity of CCPs decreased with increasing cell spreading area in a manner that was dependent on the cortical actin network. Our results point to another facet of the regulation of CCP dynamics, suggesting that CME may be modulated while cells change their mechanical state and remodel their actin cytoskeleton during various processes.

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References

Hochmuth R . Micropipette aspiration of living cells. J Biomech. 1999; 33(1):15-22. DOI: 10.1016/s0021-9290(99)00175-x. View

McBeath R, Pirone D, Nelson C, Bhadriraju K, Chen C . Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell. 2004; 6(4):483-95. DOI: 10.1016/s1534-5807(04)00075-9. View

Mettlen M, Loerke D, Yarar D, Danuser G, Schmid S . Cargo- and adaptor-specific mechanisms regulate clathrin-mediated endocytosis. J Cell Biol. 2010; 188(6):919-33. PMC: 2845073. DOI: 10.1083/jcb.200908078. View

Fogelson B, Mogilner A . Computational estimates of membrane flow and tension gradient in motile cells. PLoS One. 2014; 9(1):e84524. PMC: 3894945. DOI: 10.1371/journal.pone.0084524. View

Cocucci E, Gaudin R, Kirchhausen T . Dynamin recruitment and membrane scission at the neck of a clathrin-coated pit. Mol Biol Cell. 2014; 25(22):3595-609. PMC: 4230619. DOI: 10.1091/mbc.E14-07-1240. View

Saffarian S, Cocucci E, Kirchhausen T . Distinct dynamics of endocytic clathrin-coated pits and coated plaques. PLoS Biol. 2009; 7(9):e1000191. PMC: 2731173. DOI: 10.1371/journal.pbio.1000191. View

Tan J, Tien J, Pirone D, Gray D, Bhadriraju K, Chen C . Cells lying on a bed of microneedles: an approach to isolate mechanical force. Proc Natl Acad Sci U S A. 2003; 100(4):1484-9. PMC: 149857. DOI: 10.1073/pnas.0235407100. View

Iwadate Y, Yumura S . Molecular dynamics and forces of a motile cell simultaneously visualized by TIRF and force microscopies. Biotechniques. 2008; 44(6):739-50. DOI: 10.2144/000112752. View

Grassart A, Cheng A, Hong S, Zhang F, Zenzer N, Feng Y . Actin and dynamin2 dynamics and interplay during clathrin-mediated endocytosis. J Cell Biol. 2014; 205(5):721-35. PMC: 4050722. DOI: 10.1083/jcb.201403041. View

10.

Huang F, Khvorova A, Marshall W, Sorkin A . Analysis of clathrin-mediated endocytosis of epidermal growth factor receptor by RNA interference. J Biol Chem. 2004; 279(16):16657-61. DOI: 10.1074/jbc.C400046200. View

11.

Walani N, Torres J, Agrawal A . Endocytic proteins drive vesicle growth via instability in high membrane tension environment. Proc Natl Acad Sci U S A. 2015; 112(12):E1423-32. PMC: 4378438. DOI: 10.1073/pnas.1418491112. View

12.

Puthenveedu M, Von Zastrow M . Cargo regulates clathrin-coated pit dynamics. Cell. 2006; 127(1):113-24. DOI: 10.1016/j.cell.2006.08.035. View

13.

Scita G, Di Fiore P . The endocytic matrix. Nature. 2010; 463(7280):464-73. DOI: 10.1038/nature08910. View

14.

Taylor M, Lampe M, Merrifield C . A feedback loop between dynamin and actin recruitment during clathrin-mediated endocytosis. PLoS Biol. 2012; 10(4):e1001302. PMC: 3323523. DOI: 10.1371/journal.pbio.1001302. View

15.

McMahon H, Boucrot E . Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol. 2011; 12(8):517-33. DOI: 10.1038/nrm3151. View

16.

Gauthier N, Masters T, Sheetz M . Mechanical feedback between membrane tension and dynamics. Trends Cell Biol. 2012; 22(10):527-35. DOI: 10.1016/j.tcb.2012.07.005. View

17.

Stachowiak J, Brodsky F, Miller E . A cost-benefit analysis of the physical mechanisms of membrane curvature. Nat Cell Biol. 2013; 15(9):1019-27. PMC: 3813008. DOI: 10.1038/ncb2832. View

18.

Chen C, Mrksich M, Huang S, Whitesides G, Ingber D . Geometric control of cell life and death. Science. 1997; 276(5317):1425-8. DOI: 10.1126/science.276.5317.1425. View

19.

Lieber A, Yehudai-Resheff S, Barnhart E, Theriot J, Keren K . Membrane tension in rapidly moving cells is determined by cytoskeletal forces. Curr Biol. 2013; 23(15):1409-17. DOI: 10.1016/j.cub.2013.05.063. View

20.

Antonescu C, Aguet F, Danuser G, Schmid S . Phosphatidylinositol-(4,5)-bisphosphate regulates clathrin-coated pit initiation, stabilization, and size. Mol Biol Cell. 2011; 22(14):2588-600. PMC: 3135483. DOI: 10.1091/mbc.E11-04-0362. View