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Title: Studies on the morphology and spreading of human endothelial cells define key inter- and intramolecular interactions for talin1.
Authors: Kopp, PM
Bate, N
Hansen, TM
Brindle, NP
Praekelt, U
Debrand, E
Coleman, S
Mazzeo, D
Goult, BT
Gingras, AR
Pritchard, CA
Critchley, DR
Monkley, SJ
First Published: Sep-2010
Citation: EUR J CELL BIOL, 2010, 89 (9), pp. 661-673
Abstract: Talin binds to and activates integrins and is thought to couple them to cytoskeletal actin. However, functional studies on talin have been restricted by the fact that most cells express two talin isoforms. Here we show that human umbilical vein endothelial cells (HUVEC) express only talin1, and that talin1 knockdown inhibited focal adhesion (FA) assembly preventing the cells from maintaining a spread morphology, a phenotype that was rescued by GFP-mouse talin1. Thus HUVEC offer an ideal model system in which to conduct talin structure/function studies. Talin contains an N-terminal FERM domain (comprised of F1, F2 and F3 domains) and a C-terminal flexible rod. The F3 FERM domain binds beta-integrin tails, and mutations in F3 that inhibited integrin binding (W359A) or activation (L325R) severely compromised the ability of GFP-talin1 to rescue the talin1 knockdown phenotype despite the presence of a second integrin-binding site in the talin rod. The talin rod contains several actin-binding sites (ABS), and mutations in the C-terminal ABS that reduced actin-binding impaired talin1 function, whereas those that increased binding resulted in more stable FAs. The results show that both the N-terminal integrin and C-terminal actin-binding functions of talin are essential to cell spreading and FA assembly. Finally, mutations that relieve talin auto-inhibition resulted in the rapid and excessive production of FA, highlighting the importance of talin regulation within the cell.
DOI Link: 10.1016/j.ejcb.2010.05.003
eISSN: 1618-1298
Type: Journal Article
Appears in Collections:Published Articles, Dept. of Biochemistry

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