Originally published as Genetics Published Articles Ahead of Print on May 16, 2007.

Genetics, Vol. 176, 2201-2212, August 2007, Copyright © 2007
doi:10.1534/genetics.107.072967

A Rho-Binding Protein Kinase C-Like Activity Is Required for the Function of Protein Kinase N in Drosophila Development

Martha Betson* and Jeffrey Settleman*,1

* Massachusetts General Hospital Cancer Center and Harvard Medical School, Charlestown, Massachusetts 02129

1 Corresponding author: MGH Cancer Center, Bldg. 149, 13th St., Charlestown, MA 02129.
E-mail: settleman{at}helix.mgh.harvard.edu

The Rho GTPases interact with multiple downstream effectors to exert their biological functions, which include important roles in tissue morphogenesis during the development of multicellular organisms. Among the Rho effectors are the protein kinase N (PKN) proteins, which are protein kinase C (PKC)-like kinases that bind activated Rho GTPases. The PKN proteins are well conserved evolutionarily, but their biological role in any organism is poorly understood. We previously determined that the single Drosophila ortholog of mammalian PKN proteins, Pkn, is a Rho/Rac-binding kinase essential for Drosophila development. By performing "rescue" studies with various Pkn mutant constructs, we have defined the domains of Pkn required for its role during Drosophila development. These studies suggested that Rho, but not Rac binding is important for Pkn function in development. In addition, we determined that the kinase domain of PKC53E, a PKC family kinase, can functionally substitute for the kinase domain of Pkn during development, thereby exemplifying the evolutionary strategy of "combining" functional domains to produce proteins with distinct biological activities. Interestingly, we also identified a requirement for Pkn in wing morphogenesis, thereby revealing the first postembryonic function for Pkn.