Isoform-specific phosphorylation of axonemal dynein heavy chains

Axonemal dyneins power ciliary motility and phosphorylation of key intermediate and light chain components affects the regulation and properties of these motors in very distantly related organisms. It is also known that many axonemal dynein heavy chains are subject to this posttranslational modification although this has been little studied. Here we examine axonemal dynein heavy chains from a broad range of ciliated eukaryotes and identify phosphorylated sites embedded within various kinase recognition motifs such as those for protein kinase A, protein kinase C, and casein kinase II. Mapping these sites onto discrete heavy chain types reveals class-specific locations apparently mediated by different kinases. For example, we find that all <i>Chlamydomonas</i> α heavy chain phosphorylation sites are in an extended loop derived from AAA5 that arches over the coiled-coil buttress which in turn interacts with the microtubule-binding stalk. In contrast, most sites in the monomeric inner arm dyneins occur very close to the N-terminus and may be involved in assembly processes. In <i>Chlamydomonas</i>, the two cilia (termed <i>cis</i> and <i>trans</i>) exhibit different intrinsic beat frequencies and we identify cilium-specific phosphorylation patterns on both the α heavy chain and outer arm docking complex consistent with differential regulation of these motors in the two organelles.