Betson Martha, Settleman Jeffrey
Massachusetts General Hospital Cancer Center and Harvard Medical School, Charlestown, MA 02129, USA.
Genetics. 2007 Aug;176(4):2201-12. doi: 10.1534/genetics.107.072967. Epub 2007 May 16.
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.
Rho GTP酶与多种下游效应分子相互作用以发挥其生物学功能,这些功能在多细胞生物发育过程中的组织形态发生中起重要作用。Rho效应分子包括蛋白激酶N(PKN)蛋白,它们是与活化的Rho GTP酶结合的蛋白激酶C(PKC)样激酶。PKN蛋白在进化上高度保守,但其在任何生物体中的生物学作用却知之甚少。我们先前确定,哺乳动物PKN蛋白在果蝇中的单一直系同源物Pkn是果蝇发育所必需的Rho/Rac结合激酶。通过用各种Pkn突变体构建体进行“拯救”研究,我们确定了Pkn在果蝇发育过程中发挥作用所需的结构域。这些研究表明,对于Pkn在发育中的功能而言,Rho结合而非Rac结合很重要。此外,我们确定,PKC家族激酶PKC53E的激酶结构域在发育过程中可以在功能上替代Pkn的激酶结构域,从而例证了“组合”功能结构域以产生具有不同生物学活性的蛋白质的进化策略。有趣的是,我们还确定了Pkn在翅形态发生中的需求,从而揭示了Pkn的首个胚胎后功能。