Kinesin II and regulated intraflagellar transport of Chlamydomonas aurora protein kinase.

Abstract

The assembly and functioning of cilia and flagella depend on a complex system of traffic between the organelles and the cell body. Two types of transport into these organelles have been identified. The best characterized is constitutive: in a process termed intraflagellar transport (IFT), flagellar structural components are continuously carried into cilia and flagella on transport complexes termed IFT particles via the microtubule motor protein kinesin II. Previous studies have shown that the flagella of the unicellular green alga Chlamydomonas exhibit a second type of protein import that is regulated. During fertilization, the Chlamydomonas aurora protein kinase CALK undergoes regulated translocation from the cell body into the flagella. The motor that powers this second, regulated type of movement is unknown. Here, we have examined the cellular properties of the CALK in Chlamydomonas and used a kinesin II mutant to test the idea that the motor protein is essential for regulated translocation of proteins into flagella. We found that the CALK that is transported into flagella of wild-type gametes becomes part of a membrane-associated complex, that kinesin II is essential for the normal localization of this Chlamydomonas aurora protein kinase in unactivated gametes and that the cAMP-induced translocation of the protein kinase into flagella is disrupted in the fla10 mutants. Our results indicate that, in addition to its role in the constitutive transport of IFT particles and their cargo, kinesin II is essential for regulated translocation of proteins into flagella.