Researchers have isolated two E-cadherin mutant isoforms that are hypothesized to function differently from the isoform of the wild-type E-cadherin. An E-cadherin negative mammary carcinoma cell line was transfected with the mutant E-cadherin genes A (part a in the figure; triangles), or B (part b; triangles) or the wild-type E-cadherin gene (black circles) and compared to untransfected cells (open circles) in an aggregation assay. In this assay, cells arc first dissociated by trypsin treatment and then allowed to aggregate in solution over a period of minutes. Aggregating cells from mutants A and B are presented in panels a and b respectively. To demonstrate that the observed adhesion was cadherin mediated, the cells were pretreated with a nonspecific annbody (left panel) or a function-blocking anri-E-cadhenn monoclonal antibody (right panel).

a. Why do cells transfected with the wild-type E-cadherin gene have greater aggregation than control, untransfectecl cells?

b. From these data, what can be said about the function of mutants A and B?

c. Why does the addition of the anti-E-cadherin monoclonal antibody, but not the nonspecific antibody, block aggregation?

d. What would happen to the aggregation ability of the cells transfected with the wild-type E-cadherin gene if the assay were performed in media low in Ca⁺²?

(a)

(b)

Transcribed Image Text:

Nonspecific antibody Anti-E-cadherin 80 80 60 60 40 40 20 20 -20 -20 40 60 60 20 40 Time (min) Time (min) Aggregation (%)