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© The Rockefeller University Press, 0021-9525/2000/4/195/ $5.00
The Journal of Cell Biology, Volume 149, Number 1, April 3, 2000 195-208

Original Article

An Epidermal Plakin that Integrates Actin and Microtubule Networks at Cellular Junctions

Iakowos Karakesisogloua, Yanmin Yanga, and Elaine Fuchsa
a Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637

Correspondence to: Elaine Fuchs, Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology, The University of Chicago, 5841 S. Maryland Avenue, Room N314, Chicago, IL 60637. Tel:(773) 702-1347 Fax:(773) 702-0141 E-mail:nlipak{at}midway.uchicago.edu.

Plakins are cytoskeletal linker proteins initially thought to interact exclusively with intermediate filaments (IFs), but recently were found to associate additionally with actin and microtubule networks. Here, we report on ACF7, a mammalian orthologue of the Drosophila kakapo plakin genetically involved in epidermal–muscle adhesion and neuromuscular junctions. While ACF7/kakapo is divergent from other plakins in its IF-binding domain, it has at least one actin (Kd = 0.35 µM) and one microtubule (Kd ~6 µM) binding domain. Similar to its fly counterpart, ACF7 is expressed in the epidermis. In well spread epidermal keratinocytes, ACF7 discontinuously decorates the cytoskeleton at the cell periphery, including microtubules (MTs) and actin filaments (AFs) that are aligned in parallel converging at focal contacts. Upon calcium induction of intercellular adhesion, ACF7 and the cytoskeleton reorganize at cell–cell borders but with different kinetics from adherens junctions and desmosomes. Treatments with cytoskeletal depolymerizing drugs reveal that ACF7's cytoskeletal association is dependent upon the microtubule network, but ACF7 also appears to stabilize actin at sites where microtubules and microfilaments meet. We posit that ACF7 may function in microtubule dynamics to facilitate actin–microtubule interactions at the cell periphery and to couple the microtubule network to cellular junctions. These attributes provide a clear explanation for the kakapo mutant phenotype in flies.

Key Words: kakapo, ACF7, cytoskeleton, integrins, cell adhesion


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