Recruitment of vimentin to the cell surface by beta3 integrin and plectin mediates adhesion strength.
Much effort has been expended on analyzing how microfilament and microtubule cytoskeletons dictate the interaction of cells with matrix at adhesive sites called focal adhesions (FAs). However, vimentin intermediate filaments (IFs) also associate with the cell surface at FAs in endothelial cells. Here, we show that IF recruitment to FAs in endothelial cells requires beta3 integrin, plectin and the microtubule cytoskeleton, and is dependent on microtubule motors. In CHO cells, which lack beta3 integrin but contain vimentin, IFs appear to be collapsed around the nucleus, whereas in CHO cells expressing beta3 integrin (CHOwtbeta3), vimentin IFs extend to FAs at the cell periphery. This recruitment is regulated by tyrosine residues in the beta3 integrin cytoplasmic tail. Moreover, CHOwtbeta3 cells exhibit significantly greater adhesive strength than CHO or CHO cells expressing mutated beta3 integrin proteins. These differences require an intact vimentin network. Therefore, vimentin IF recruitment to the cell surface is tightly regulated and modulates the strength of adhesion of cells to their substrate.
Bhattacharya
Ramona
R
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.
Gonzalez
Annette M
AM
Debiase
Phillip J
PJ
Trejo
Humberto E
HE
Goldman
Robert D
RD
Flitney
Frederick W
FW
Jones
Jonathan C R
JC
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003599:Cytoskeleton / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism*; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D007382:Intermediate Filaments / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000378:metabolism; D051419:Paxillin / Q000235:genetics / Q000378:metabolism; D051190:Plectin / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014746:Vimentin / Q000235:genetics / Q000378:metabolism*
D039661:Integrin beta3; D020409:Molecular Motor Proteins; D051419:Paxillin; D051190:Plectin; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D014746:Vimentin; D051416:Focal Adhesion Protein-Tyrosine Kinases