title: The DPY-30 domain and its flanking sequence mediate the assembly and modulation of flagellar radial spoke complexes.
abstract: RIIa is known as the dimerization and docking (D/D) domain of the cyclic AMP (cAMP)-dependent protein kinase. However, numerous molecules, including radial spoke protein 2 (RSP2) in Chlamydomonas flagella, also contain an RIIa or a similar DPY-30 domain. To elucidate new roles of D/D domain-containing proteins, we investigated a panel of RSP2 mutants. An RSP2 mutant had paralyzed flagella defective in RSP2 and multiple subunits near the spokehead. New transgenic strains lacking only the DPY-30 domain in RSP2 were also paralyzed. In contrast, motility was restored in strains that lacked only RSP2's calmodulin-binding C-terminal region. These cells swam normally in dim light but could not maintain typical swimming trajectories under bright illumination. In both deletion transgenic strains, the subunits near the spokehead were restored, but their firm attachment to the spokestalk required the DPY-30 domain. We postulate that the DPY-30-helix dimer is a conserved two-prong linker, required for normal motility, organizing duplicated subunits in the radial spoke stalk and formation of a symmetrical spokehead. Further, the dispensable calmodulin-binding region appears to fine-tune the spokehead for regulation of "steering" motility in the green algae. Thus, in general, D/D domains may function to localize molecular modules for both the assembly and modulation of macromolecular complexes.
authors.affiliation: Department of Biological Sciences, Marquette University, Milwaukee, WI, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Gopal
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Foster
authors.forename: Pinfen
authors.initials: P
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002465:Cell Movement; D002696:Chlamydomonas / Q000737:chemistry* / Q000166:cytology* / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D005407:Flagella / Q000737:chemistry* / Q000502:physiology*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D010940:Plant Proteins; D017868:Cyclic AMP-Dependent Protein Kinases