Control of ciliary orientation through cAMP-dependent phosphorylation of axonemal proteins inParamecium caudatum

Abstract
Ciliary reorientations in response to cAMP do not take place after a brief digestion with trypsin in ciliated cortical sheets from Triton‐glycerol‐extracted Paramecium. In this study, we examined the effects of tryptic digestion on the cAMP‐dependent phosphorylation of axonemal proteins to clarify the relationship between phosphorylation and ciliary reorientation. As reported for Paramecium tetraurelia, cAMP stimulated phosphorylations of the 29 kDa and 65 kDa axonemal polypeptides also in Paramecium caudatum. After a brief digestion of axonemes by trypsin, none of the cAMP‐dependent phosphorylations occurred. On the other hand, the 29 kDa polypeptide still remained to be labeled after a brief digestion of axonemes that had previously been labeled with 32P in the presence of cAMP, which indicates that this brief digestion breaks down endogenous cAMP‐dependent protein kinases but not phosphorylated proteins. This must be the reason that trypsin‐treated cilia on the sheets cannot reorient towards the posterior part of the cell. Our results indicate that cAMP regulates not only the beat frequency but also the ciliary orientation via phosphorylation of dynein subunits in Paramecium. Cell Motil. Cytoskeleton 45:263–271, 2000