Calcium Levels Affect the Ability to Immunolocalize Calmodulin to Cortical Microtubules

Calcium affects the stability of cortical microtubules (MTs) in lysed protoplasts. This calmodulin (CaM)-mediated interaction may provide a mechanism that serves to integrate cellular behavior with MT function. To test the hypothesis that CaM associates with these MTs, monoclonal antibodies were produced against CaM, and one (designated mAb1D10) was selected for its suitability as an immunocytochemical reagent. It is shown that CaM associates with the cortical MTs of cultured carrot (Daucus carota L.) and tobacco (Nicotiana tabacum L.) cells. Inasmuch as CaM interacts with calcium and affects the behavior of these MTs, we hypothesized that calcium would alter this association. To test this, protoplasts containing taxol-stabilized MTs were lysed in the presence of various concentrations of calcium and examined for the association of CaM with cortical MTs. At 1 [mu]M calcium, many protoplasts did not have CaM in association with the cortical MTs, whereas at 3.6 [mu]M calcium, this association was completely abolished. Control experiments were performed to eliminate alternate explanations including differential antibody binding in the presence of calcium and/or taxol, detergent-induced redistribution of antigen, and epitope masking. The results are discussed in terms of a model in which CaM associates with MTs via two types of interactions, one that occurs in the presence of calcium and another that occurs only in its absence.