A CDK-related kinase regulates the length and assembly of flagella in Chlamydomonas.
Little is known about how cells regulate the size of their organelles. In this study, we find that proper flagellar length control in Chlamydomonas reinhardtii requires the activity of a new member of the cyclin-dependent kinase (CDK) family, which is encoded by the LF2 (long flagella 2) gene. This novel CDK contains all of the important residues that are essential for kinase activity but lacks the cyclin-binding motif PSTAIRE. Analysis of genetic lesions in a series of lf2 mutant alleles and site-directed mutagenesis of LF2p reveals that improper flagellar length and defective flagellar assembly correlate with the extent of disruption of conserved kinase structures or residues by mutations. LF2p appears to interact with both LF1p and LF3p in the cytoplasm, as indicated by immunofluorescence localization, sucrose density gradients, cell fractionation, and yeast two-hybrid experiments. We propose that LF2p is the catalytic subunit of a regulatory kinase complex that controls flagellar length and flagellar assembly.
Tam
Lai-Wa
LW
Department of Plant Biology, University of Minnesota, St. Paul, MN 55108, USA.
Wilson
Nedra F
NF
Lefebvre
Paul A
PA
D000483:Alleles; D020816:Amino Acid Motifs; D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000201:enzymology / Q000648:ultrastructure*; D018844:Cyclin-Dependent Kinases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D005407:Flagella / Q000201:enzymology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D020798:Two-Hybrid System Techniques
D015800:Protozoan Proteins; D012333:RNA, Messenger; D018844:Cyclin-Dependent Kinases