DOI: 10.1091/mbc.e02-03-0142. RACK1 Regulates Integrin-mediated Adhesion, Protrusion, and Chemotactic Cell Migration via Its Src-binding Site

RACK1 Regulates Integrin-mediated Adhesion, Protrusion, and Chemotactic Cell Migration via Its Src-binding Site

10.1091/mbc.e02-03-0142

Crossref journal-article

American Society for Cell Biology (ASCB)

Molecular Biology of the Cell (1076)

Abstract

Mammalian cDNA expression cloning was used to identify novel regulators of integrin-mediated cell-substratum adhesions. Using a focal adhesion morphology screen, we identified a cDNA with homology to a receptor for activated protein kinase C (RACK1) that induced a loss of central focal adhesions and stress fibers in CHO-K1 cells. The identified cDNA was a C-terminal truncated form of RACK1 that had one of the putative protein kinase C binding sites but lacked the region proposed to bind the β integrin cytoplasmic domain and the tyrosine kinase Src. To investigate the role of RACK1 during cell spreading and migration, we tagged RACK1, a C-terminal truncated RACK1 and a point mutant that does not bind Src (RACK Y246F) with green fluorescent protein and expressed them in CHO-K1 cells. We found that RACK1 regulates the organization of focal adhesions and that it localizes to a subset of nascent focal complexes in areas of protrusion that contain paxillin but not vinculin. We also found that RACK1 regulates cell protrusion and chemotactic migration through its Src binding site. Together, these findings suggest that RACK1 regulates adhesion, protrusion, and chemotactic migration through its interaction with Src.

Bibliography

Cox, E. A., Bennin, D., Doan, A. T., OâToole, T., & Huttenlocher, A. (2003). RACK1 Regulates Integrin-mediated Adhesion, Protrusion, and Chemotactic Cell Migration via Its Src-binding Site. Molecular Biology of the Cell, 14(2), 658â669.

Authors 5

Elisabeth A. Cox (first)
David Bennin (additional)
Ashley T. Doan (additional)
Timothy O'Toole (additional)
Anna Huttenlocher (additional)

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Dates

Type	When
Created	22 years, 6 months ago (Feb. 14, 2003, 7:02 p.m.)
Deposited	4 years, 2 months ago (June 4, 2021, 2:13 p.m.)
Indexed	2 months ago (June 26, 2025, 9:09 a.m.)
Issued	22 years, 7 months ago (Feb. 1, 2003)
Published	22 years, 7 months ago (Feb. 1, 2003)
Published Print	22 years, 7 months ago (Feb. 1, 2003)

Funders 0

None

BibTeX

@article{Cox_2003, title={RACK1 Regulates Integrin-mediated Adhesion, Protrusion, and Chemotactic Cell Migration via Its Src-binding Site}, volume={14}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e02-03-0142}, DOI={10.1091/mbc.e02-03-0142}, number={2}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Cox, Elisabeth A. and Bennin, David and Doan, Ashley T. and O’Toole, Timothy and Huttenlocher, Anna}, editor={Schwartz, Martin}, year={2003}, month=feb, pages={658–669} }

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  "abstract": "<jats:p>Mammalian cDNA expression cloning was used to identify novel regulators of integrin-mediated cell-substratum adhesions. Using a focal adhesion morphology screen, we identified a cDNA with homology to a receptor for activated protein kinase C (RACK1) that induced a loss of central focal adhesions and stress fibers in CHO-K1 cells. The identified cDNA was a C-terminal truncated form of RACK1 that had one of the putative protein kinase C binding sites but lacked the region proposed to bind the \u03b2 integrin cytoplasmic domain and the tyrosine kinase Src. To investigate the role of RACK1 during cell spreading and migration, we tagged RACK1, a C-terminal truncated RACK1 and a point mutant that does not bind Src (RACK Y246F) with green fluorescent protein and expressed them in CHO-K1 cells. We found that RACK1 regulates the organization of focal adhesions and that it localizes to a subset of nascent focal complexes in areas of protrusion that contain paxillin but not vinculin. We also found that RACK1 regulates cell protrusion and chemotactic migration through its Src binding site. Together, these findings suggest that RACK1 regulates adhesion, protrusion, and chemotactic migration through its interaction with Src.</jats:p>",
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