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Studies on the morphology and spreading of human endothelial cells define key inter- and intramolecular interactions for talin1.

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 - Eur. J. Cell Biol. (2010)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Leicester, Leicester, UK.

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Related in: MedlinePlus

Use of HUVEC for talin1 knockdown. (A) Quantitative RT-PCR of talin1/talin2 expression in either hFF or HUVEC. (B) Western blot of cell lysate from hFFs or HUVECs transfected with talin1 siRNA for 72 h. Tubulin was loading control. (C) Phase micrographs of HUVEC either untransfected or transfected with conRNA or talin1 siRNA for 72 h and replated onto glass for 24 h. (D) HUVEC transfected with either a talin1 siRNA or conRNA were replated on glass coverslips 72 h post transfection. Brightfield images were recorded at the times indicated. Scale bars, 10 μm. For talin 1 siRNA transfected cells, 3 out of the 6 cells shown were able to adhere and spread initially, but they started to arborize after 150 min or could not maintain spreading. Three cells failed to spread at all.
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d32e1511: Use of HUVEC for talin1 knockdown. (A) Quantitative RT-PCR of talin1/talin2 expression in either hFF or HUVEC. (B) Western blot of cell lysate from hFFs or HUVECs transfected with talin1 siRNA for 72 h. Tubulin was loading control. (C) Phase micrographs of HUVEC either untransfected or transfected with conRNA or talin1 siRNA for 72 h and replated onto glass for 24 h. (D) HUVEC transfected with either a talin1 siRNA or conRNA were replated on glass coverslips 72 h post transfection. Brightfield images were recorded at the times indicated. Scale bars, 10 μm. For talin 1 siRNA transfected cells, 3 out of the 6 cells shown were able to adhere and spread initially, but they started to arborize after 150 min or could not maintain spreading. Three cells failed to spread at all.


Studies on the morphology and spreading of human endothelial cells define key inter- and intramolecular interactions for talin1.

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 - Eur. J. Cell Biol. (2010)

Use of HUVEC for talin1 knockdown. (A) Quantitative RT-PCR of talin1/talin2 expression in either hFF or HUVEC. (B) Western blot of cell lysate from hFFs or HUVECs transfected with talin1 siRNA for 72 h. Tubulin was loading control. (C) Phase micrographs of HUVEC either untransfected or transfected with conRNA or talin1 siRNA for 72 h and replated onto glass for 24 h. (D) HUVEC transfected with either a talin1 siRNA or conRNA were replated on glass coverslips 72 h post transfection. Brightfield images were recorded at the times indicated. Scale bars, 10 μm. For talin 1 siRNA transfected cells, 3 out of the 6 cells shown were able to adhere and spread initially, but they started to arborize after 150 min or could not maintain spreading. Three cells failed to spread at all.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2958305&req=5

d32e1511: Use of HUVEC for talin1 knockdown. (A) Quantitative RT-PCR of talin1/talin2 expression in either hFF or HUVEC. (B) Western blot of cell lysate from hFFs or HUVECs transfected with talin1 siRNA for 72 h. Tubulin was loading control. (C) Phase micrographs of HUVEC either untransfected or transfected with conRNA or talin1 siRNA for 72 h and replated onto glass for 24 h. (D) HUVEC transfected with either a talin1 siRNA or conRNA were replated on glass coverslips 72 h post transfection. Brightfield images were recorded at the times indicated. Scale bars, 10 μm. For talin 1 siRNA transfected cells, 3 out of the 6 cells shown were able to adhere and spread initially, but they started to arborize after 150 min or could not maintain spreading. Three cells failed to spread at all.
Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Leicester, Leicester, UK.

Show MeSH
Related in: MedlinePlus