The LC7 light chains of Chlamydomonas flagellar dyneins interact with components required for both motor assembly and regulation.
Members of the LC7/Roadblock family of light chains (LCs) have been found in both cytoplasmic and axonemal dyneins. LC7a was originally identified within Chlamydomonas outer arm dynein and associates with this motor's cargo-binding region. We describe here a novel member of this protein family, termed LC7b that is also present in the Chlamydomonas flagellum. Levels of LC7b are reduced approximately 20% in axonemes isolated from strains lacking inner arm I1 and are approximately 80% lower in the absence of the outer arms. When both dyneins are missing, LC7b levels are diminished to <10%. In oda9 axonemal extracts that completely lack outer arms, LC7b copurifies with inner arm I1, whereas in ida1 extracts that are devoid of I1 inner arms it associates with outer arm dynein. We also have observed that some LC7a is present in both isolated axonemes and purified 18S dynein from oda1, suggesting that it is also a component of both the outer arm and inner arm I1. Intriguingly, in axonemal extracts from the LC7a null mutant, oda15, which assembles approximately 30% of its outer arms, LC7b fails to copurify with either dynein, suggesting that it interacts with LC7a. Furthermore, both the outer arm gamma heavy chain and DC2 from the outer arm docking complex completely dissociate after salt extraction from oda15 axonemes. EDC cross-linking of purified dynein revealed that LC7b interacts with LC3, an outer dynein arm thioredoxin; DC2, an outer arm docking complex component; and also with the phosphoprotein IC138 from inner arm I1. These data suggest that LC7a stabilizes both the outer arms and inner arm I1 and that both LC7a and LC7b are involved in multiple intradynein interactions within both dyneins.
DiBella
Linda M
LM
Department of Biochemistry, University of Connecticut Health Center, Farmington, CT 06030-3305, USA.
Sakato
Miho
M
Patel-King
Ramila S
RS
Pazour
Gregory J
GJ
King
Stephen M
SM
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000166:cytology / Q000378:metabolism*; D004398:Dyneins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D020409:Molecular Motor Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D015800:Protozoan Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D002352:Carrier Proteins; C492351:LC7 protein, Chlamydomonas; D020409:Molecular Motor Proteins; D010940:Plant Proteins; D020033:Protein Isoforms; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D004398:Dyneins