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Dynamics of focal adhesion kinase and paxillin in lamellipodial protrusion of migrating endothelial cells
Author(s) -
Hu Ying-Li,
Lu Shaoying,
Wang Yingxiao,
Chien Shu
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1129.13
Subject(s) - paxillin , focal adhesion , microbiology and biotechnology , ptk2 , cell migration , pseudopodia , actin , transfection , chemistry , cell adhesion , cell , biology , kinase , phosphorylation , protein kinase a , biochemistry , mitogen activated protein kinase kinase , gene
The dynamics of focal adhesions (FAs) can regulate cell migration, intracellular signaling, and remodeling of actin filaments. FA turnover is dependent on focal adhesion kinase (FAK) and paxillin, which are involved in FA disassembly and formation. We used time‐lapse double‐color imaging to monitor concurrently the spatial‐temporal dynamics of FAK and paxillin in live endothelial cells (ECs) co‐transfected with GFP‐FAK and DsRed‐paxillin. In comparison to fluorescence intensity (FI) of paxillin‐FAs, FI of FAK‐FAs was markedly higher at cell front (lamellipodial protrusion), approximately equal at cell center, and lower at cell rear. Consistently, the mean FAK/Paxillin FI ratio from multiple cells is highest at cell front (mean±SE: 4.73±0.48), almost equal to 1 at cell center (0.95±0.11), and lowest at cell rear (0.64±0.05). In ECs co‐transfected with RFP‐actin and GFP‐FAK, FAK is found to be associated with actin filaments at cell leading edge, but paxillin shows minimal presence. Our results suggest that FAK‐FAs are assembled in lamellipodial protrusion earlier than paxillin. While both FAK and paxillin play important roles in modulating FA dynamics during cell migration, the FAK associated with actin filaments promotes FA formation ahead of paxillin at the cell leading edge. This work was supported by NHLBI Research Grants HL‐104402 and HL‐106579 (S.C.).