Epithelial and fibroblastoid cells contain numerous cell-type specific putative microtubule-regulating proteins, among which are ezrin and fodrin.
Upon cell junction formation, the microtubules of polarizing epithelial cells become reorganized by unknown signaling mechanisms and regulating proteins. Microtubule-associated (MAPs) and other types of proteins are likely to be involved in this process, but most of these are unknown. Such proteins are called here collectively microtubule-regulating proteins (MRPs). As a first step towards their characterization, we used co-sedimentation of cytosolic proteins of MDCK cells and A72, a dog fibroblastoid line, with an excess of taxol-stabilized MTs, to obtain a cell fraction enriched in putative MRPs ("MRPs"). Additional tests have led to the inventory of around 40 "MRPs" among the 80 proteins present in the microtubule pellet. We also found that "MRPs" are recovered in higher amounts from MDCK cytosol, and that half of these are cell-type specific. These results corroborate data from yeast cells and insect eggs, and show that in mammalian somatic cells too, a large number of proteins seems to be involved in microtubule regulation, and that different cell types use a specific set of MRPs. "MRPs" found in both cell types are the intermediate chain of cytoplasmic dynein, Arp1, the major subunit of the dynactin complex, and CLIP-170. Two MDCK-specific "MRPs" were identified as the actin-binding proteins ezrin and alpha-fodrin. These results are discussed with regard to a possible involvement of ezrin and fodrin in morphogenetic interactions of microtubules with the membrane cytoskeleton in polarizing epithelia upon junction formation.
Shestakova
E
E
Institut Jacques Monod, Department of Supramolecular and Cellular Biology, Université Paris VII, France.
Vandekerckhove
J
J
De Mey
J R
JR
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D002460:Cell Line; D003598:Cytoskeletal Proteins; D004285:Dogs; D004847:Epithelial Cells / Q000737:chemistry*; D005347:Fibroblasts / Q000737:chemistry*; D007668:Kidney / Q000166:cytology; D008840:Microfilament Proteins / Q000502:physiology*; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010750:Phosphoproteins / Q000502:physiology*; D011485:Protein Binding; D013049:Spectrin; D000099149:Ezrin
D000199:Actins; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D013049:Spectrin; D000099149:Ezrin