Stimulation of in vitro motility of Chlamydomonas axonemes by inhibition of cAMP-dependent phosphorylation.
When demembranated axonemes of Chlamydomonas were reactivated with Mg-ATP, the proportion of motile axonemes was significantly increased by the presence of either phosphodiesterase (PDE) or protein inhibitor of cAMP-dependent kinase (PKI). The effect of PDE was cancelled by the addition of cAMP. These findings strongly suggest that the axoneme samples have endogenous cAMP, which can reduce the proportion of motile axonemes via phosphorylation. This inhibitory effect of cAMP on Chlamydomonas axonemes is opposite to its stimulatory effect on the axonemal motility in other organisms so far reported. PKI or PDE activated the motility either in the absence of Ca2+, when the axonemes beat with an asymmetric waveform, or in 10(-5) M Ca2+, when the axonemes beat with a symmetric waveform. This cAMP-dependent regulation of motility was observed with the axonemes from which detergent-soluble material had been removed, indicating that the proteins responsible for the regulation still remained in the axonemes. Preliminary in vitro phosphorylation studies have implicated two polypeptides as candidates for the target protein of cAMP-dependent protein kinase: one with a molecular weight of 270 kD and the other with a much larger molecular weight.
Hasegawa
E
E
Institute of Molecular Biology, Faculty of Science, Nagoya University, Japan.
Hayashi
H
H
Asakura
S
S
Kamiya
R
R
D002118:Calcium / Q000502:physiology; D002147:Calmodulin / Q000494:pharmacology; D002465:Cell Movement; D002696:Chlamydomonas / Q000648:ultrastructure; D000242:Cyclic AMP / Q000502:physiology*; D005407:Flagella / Q000502:physiology*; D066298:In Vitro Techniques; D010750:Phosphoproteins / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors
D002147:Calmodulin; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases; D002118:Calcium