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J Biol Chem, Vol. 274, Issue 21, 15085-15094, May 21, 1999

Calreticulin Affects Focal Contact-dependent but Not Close Contact-dependent Cell-substratum Adhesion

Marc P. Fadel, Ewa Dziak, Chun-Min Lo^§, Jack Ferrier^§, Nasrin Mesaeli^¶, Marek Michalak^¶, and Michal Opas

From the  Department of Anatomy and Cell Biology, University of Toronto, Toronto, Ontario M5S 1A8, Canada, the ^§ Medical Research Council Group in Periodontal Physiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada, and the ^¶ Medical Research Council Group in Molecular Biology of Membranes, Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada

We used two cell lines expressing fast (RPE_fast) and slow (RPE_slow) attachment kinetics to investigate mechanisms of cell-substratum adhesion. We show that the abundance of a cytoskeletal protein, vinculin, is dramatically decreased in RPE_fast 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 RPE_fast cells. After RPE_fast 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. RPE_fast 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 RPE_fast cells lacked extensive focal contacts and adhered weakly but attached fast to the substratum. In contrast, the RPE_slow cells that expressed calreticulin and vinculin abundantly developed numerous and prominent focal contacts slowly, but adhered strongly. Thus, while the calreticulin overexpressing RPE_slow cells "grip" the substratum with focal contacts, calreticulin underexpressing RPE_fast cells use close contacts to "stick" to it.

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