TY - JOUR
T1 - Differences in elasticity of vinculin-deficient F9 cells measured by magnetometry and atomic force microscopy
AU - Goldmann, Wolfgang H.
AU - Galneder, Reinhard
AU - Ludwig, Markus
AU - Xu, Weiming
AU - Adamson, Eileen D.
AU - Wang, Ning
AU - Ezzell, Robert M.
N1 - We thank Dr. Manfred Radmacher for helpful comments and Judith Feldmann for proof-reading the manuscript. This work was supported by the American Cancer Society, NASA NAG5-4839, NIH HL-33009, Deutsche Forschungsgemeinschaft Go 598/3-1, and NATO (CRG 970205). Excerpts of this work were presented at the annual ASCB Meeting (1996) in San Francisco as a poster and published in abstract form in Mol. Biol. Cell (1996) 7S, 385a.
PY - 1998/3/15
Y1 - 1998/3/15
N2 - We have investigated a mouse F9 embryonic carcinoma cell line, in which both vinculin genes were inactivated by homologous recombination, that exhibits defective adhesion and spreading [Coll et al. (1995) Proc. Natl. Acad. Sci. USA 92, 9161-9165]. Using a magnetometer and RGD-coated magnetic microbeads, we measured the local effect of loss and replacement of vinculin on mechanical force transfer across integrins. Vinculin-deficient F9Vin(-/-) cells showed a 21% difference in relative stiffness compared to wild-type cells. This was restored to near wild-type levels after transfection and constitutive expression of increasing amounts of vinculin into F9Vin(-/-) cells. In contrast, the transfection of vinculin constructs deficient in amino acids 1-288 (containing the talin- and α-actinin-binding site) or substituting tyrosine for phenylalanine (phosphorylation site, amino acid 822) in F9Vin(-/-) cells resulted in partial restoration of stiffness. Using atomic force microscopy to map the relative elasticity of entire F9 cells by 128 x 128 (n = 16,384) force scans, we observed a correlation with magnetometer measurements. These findings suggest that vinculin may promote cell adhesion and spreading by stabilizing focal adhesions and transferring mechanical stresses that drive cytoskeletal remodeling, thereby affecting the elastic properties of the cell.
AB - We have investigated a mouse F9 embryonic carcinoma cell line, in which both vinculin genes were inactivated by homologous recombination, that exhibits defective adhesion and spreading [Coll et al. (1995) Proc. Natl. Acad. Sci. USA 92, 9161-9165]. Using a magnetometer and RGD-coated magnetic microbeads, we measured the local effect of loss and replacement of vinculin on mechanical force transfer across integrins. Vinculin-deficient F9Vin(-/-) cells showed a 21% difference in relative stiffness compared to wild-type cells. This was restored to near wild-type levels after transfection and constitutive expression of increasing amounts of vinculin into F9Vin(-/-) cells. In contrast, the transfection of vinculin constructs deficient in amino acids 1-288 (containing the talin- and α-actinin-binding site) or substituting tyrosine for phenylalanine (phosphorylation site, amino acid 822) in F9Vin(-/-) cells resulted in partial restoration of stiffness. Using atomic force microscopy to map the relative elasticity of entire F9 cells by 128 x 128 (n = 16,384) force scans, we observed a correlation with magnetometer measurements. These findings suggest that vinculin may promote cell adhesion and spreading by stabilizing focal adhesions and transferring mechanical stresses that drive cytoskeletal remodeling, thereby affecting the elastic properties of the cell.
KW - Vinculin-regulated cellular elasticity
UR - https://www.scopus.com/pages/publications/0031817205
UR - https://www.scopus.com/pages/publications/0031817205#tab=citedBy
U2 - 10.1006/excr.1997.3915
DO - 10.1006/excr.1997.3915
M3 - Article
C2 - 9521841
AN - SCOPUS:0031817205
SN - 0014-4827
VL - 239
SP - 235
EP - 242
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -