Calreticulin affects focal contact-dependent but not close contact-dependent cell-substratum adhesion.
We used two cell lines expressing fast (RPEfast) and slow (RPEslow) attachment kinetics to investigate mechanisms of cell-substratum adhesion. We show that the abundance of a cytoskeletal protein, vinculin, is dramatically decreased in RPEfast cells. This coincides with the diminished expression level of an endoplasmic reticulum chaperone, calreticulin. Both protein and mRNA levels for calreticulin and vinculin were decreased in RPEfast cells. After RPEfast cells were transfected with cDNA encoding calreticulin, both the expression of endoplasmic reticulum-resident calreticulin and cytoplasmic vinculin increased. The abundance of other adhesion-related proteins was not affected. RPEfast cells underexpressing calreticulin displayed a dramatic increase in the abundance of total cellular phosphotyrosine suggesting that the effects of calreticulin on cell adhesiveness may involve modulation of the activities of protein tyrosine kinases or phosphatases which may affect the stability of focal contacts. The calreticulin and vinculin underexpressing RPEfast cells lacked extensive focal contacts and adhered weakly but attached fast to the substratum. In contrast, the RPEslow cells that expressed calreticulin and vinculin abundantly developed numerous and prominent focal contacts slowly, but adhered strongly. Thus, while the calreticulin overexpressing RPEslow cells "grip" the substratum with focal contacts, calreticulin underexpressing RPEfast cells use close contacts to "stick" to it.
D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000502:physiology*; D037282:Calreticulin; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D005136:Eye Proteins / Q000235:genetics / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000502:physiology*; D010641:Phenotype; D019000:Phosphotyrosine / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology; D043562:Receptors, G-Protein-Coupled; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000502:physiology*; D016596:Vinculin / Q000502:physiology