title: The ciliary inner dynein arm, I1 dynein, is assembled in the cytoplasm and transported by IFT before axonemal docking.
abstract: To determine mechanisms of assembly of ciliary dyneins, we focused on the Chlamydomonas inner dynein arm, I1 dynein, also known as dynein f. I1 dynein assembles in the cytoplasm as a 20S complex similar to the 20S I1 dynein complex isolated from the axoneme. The intermediate chain subunit, IC140 (IDA7), and heavy chains (IDA1, IDA2) are required for 20S I1 dynein preassembly in the cytoplasm. Unlike I1 dynein derived from the axoneme, the cytoplasmic 20S I1 complex will not rebind I1-deficient axonemes in vitro. To test the hypothesis that I1 dynein is transported to the distal tip of the cilia for assembly in the axoneme, we performed cytoplasmic complementation in dikaryons formed between wild-type and I1 dynein mutant cells. Rescue of I1 dynein assembly in mutant cilia occurred first at the distal tip and then proceeded toward the proximal axoneme. Notably, in contrast to other combinations, I1 dynein assembly was significantly delayed in dikaryons formed between ida7 and ida3. Furthermore, rescue of I1 dynein assembly required new protein synthesis in the ida7 × ida3 dikaryons. On the basis of the additional observations, we postulate that IDA3 is required for 20S I1 dynein transport. Cytoplasmic complementation in dikaryons using the conditional kinesin-2 mutant, fla10-1 revealed that transport of I1 dynein is dependent on kinesin-2 activity. Thus, I1 dynein complex assembly depends upon IFT for transport to the ciliary distal tip prior to docking in the axoneme.
authors.affiliation: Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia.
authors.forename: Rasagnya
authors.initials: R
authors.lastname: Viswanadha
authors.forename: Emily L
authors.initials: EL
authors.lastname: Hunter
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Yamamoto
authors.forename: Maureen
authors.initials: M
authors.lastname: Wirschell
authors.forename: Lea M
authors.initials: LM
authors.lastname: Alford
authors.forename: Susan K
authors.initials: SK
authors.lastname: Dutcher
authors.forename: Winfield S
authors.initials: WS
authors.lastname: Sale
mesh_terms: D054468:Axoneme / Q000378:metabolism*; D001692:Biological Transport; D002696:Chlamydomonas / Q000378:metabolism*; D002923:Cilia / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D010940:Plant Proteins / Q000378:metabolism; D014176:Protein Biosynthesis
chemical_list: D010940:Plant Proteins; D004398:Dyneins; D016547:Kinesins
keywords: Chlamydomonas dikaryon zygotes; I1 dynein; axonemes; cilia; flagella