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The Mr 140,000 intermediate chain of Chlamydomonas flagellar inner arm dynein is a WD-repeat protein implicated in dynein arm anchoring.
- Source :
-
Molecular biology of the cell [Mol Biol Cell] 1998 Dec; Vol. 9 (12), pp. 3335-49. - Publication Year :
- 1998
-
Abstract
- Previous structural and biochemical studies have revealed that the inner arm dynein I1 is targeted and anchored to a unique site located proximal to the first radial spoke in each 96-nm axoneme repeat on flagellar doublet microtubules. To determine whether intermediate chains mediate the positioning and docking of dynein complexes, we cloned and characterized the 140-kDa intermediate chain (IC140) of the I1 complex. Sequence and secondary structural analysis, with particular emphasis on beta-sheet organization, predicted that IC140 contains seven WD repeats. Reexamination of other members of the dynein intermediate chain family of WD proteins indicated that these polypeptides also bear seven WD/beta-sheet repeats arranged in the same pattern along each intermediate chain protein. A polyclonal antibody was raised against a 53-kDa fusion protein derived from the C-terminal third of IC140. The antibody is highly specific for IC140 and does not bind to other dynein intermediate chains or proteins in Chlamydomonas flagella. Immunofluorescent microscopy of Chlamydomonas cells confirmed that IC140 is distributed along the length of both flagellar axonemes. In vitro reconstitution experiments demonstrated that the 53-kDa C-terminal fusion protein binds specifically to axonemes lacking the I1 complex. Chemical cross-linking indicated that IC140 is closely associated with a second intermediate chain in the I1 complex. These data suggest that IC140 contains domains responsible for the assembly and docking of the I1 complex to the doublet microtubule cargo.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Chlamydomonas genetics
Cloning, Molecular
DNA Primers genetics
DNA, Protozoan genetics
Dyneins genetics
Flagella enzymology
Genes, Protozoan
Molecular Sequence Data
Molecular Weight
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Repetitive Sequences, Amino Acid
Sequence Homology, Amino Acid
Chlamydomonas enzymology
Dyneins chemistry
Dyneins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1059-1524
- Volume :
- 9
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Molecular biology of the cell
- Publication Type :
- Academic Journal
- Accession number :
- 9843573
- Full Text :
- https://doi.org/10.1091/mbc.9.12.3335