Article ; Online: R-Ras regulates migration through an interaction with filamin A in melanoma cells.
PloS one
2010 Volume 5, Issue 6, Page(s) e11269
Abstract: ... in the initiation and progression of metastasis. R-Ras is one of several small GTPases that regulate cell adhesion ... Using a yeast two-hybrid approach we sought to identify novel R-Ras binding proteins that might mediate ... require FLNa for motility. We further show that R-Ras and FLNa interact in co-immunoprecipitations and ...
| Abstract | Background: Changes in cell adhesion and migration in the tumor microenvironment are key in the initiation and progression of metastasis. R-Ras is one of several small GTPases that regulate cell adhesion and migration on the extracellular matrix, however the mechanism has not been completely elucidated. Using a yeast two-hybrid approach we sought to identify novel R-Ras binding proteins that might mediate its effects on integrins. Methods and findings: We identified Filamin A (FLNa) as a candidate interacting protein. FLNa is an actin-binding scaffold protein that also binds to integrin beta1, beta2 and beta7 tails and is associated with diverse cell processes including cell migration. Indeed, M2 melanoma cells require FLNa for motility. We further show that R-Ras and FLNa interact in co-immunoprecipitations and pull-down assays. Deletion of FLNa repeat 3 (FLNaDelta3) abrogated this interaction. In M2 melanoma cells active R-Ras co-localized with FLNa but did not co-localize with FLNa lacking repeat 3. Thus, activated R-Ras binds repeat 3 of FLNa. The functional consequence of this interaction was that active R-Ras and FLNa coordinately increased cell migration. In contrast, co-expression of R-Ras and FLNaDelta3 had a significantly reduced effect on migration. While there was enhancement of integrin activation and fibronectin matrix assembly, cell adhesion was not altered. Finally, siRNA knockdown of endogenous R-Ras impaired FLNa-dependent fibronectin matrix assembly. Conclusions: These data support a model in which R-Ras functionally associates with FLNa and thereby regulates integrin-dependent migration. Thus in melanoma cells R-Ras and FLNa may cooperatively promote metastasis by enhancing cell migration. |
|---|---|
| MeSH term(s) | Base Sequence ; Blotting, Western ; Cell Movement/physiology ; Contractile Proteins/physiology ; DNA Primers ; Filamins ; Humans ; Melanoma/pathology ; Microfilament Proteins/physiology ; Microscopy, Fluorescence ; ras Proteins/physiology |
| Chemical Substances | Contractile Proteins ; DNA Primers ; Filamins ; Microfilament Proteins ; RRAS protein, human (EC 3.6.1.-) ; ras Proteins (EC 3.6.5.2) |
| Language | English |
| Publishing date | 2010-06-23 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. |
| ISSN | 1932-6203 |
| ISSN (online) | 1932-6203 |
| DOI | 10.1371/journal.pone.0011269 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
More links
Kategorien
Order via subito
This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.
Inter-library loan at ZB MED
Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.