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Department of Anatomy and Cell Biology, SUNY Health Science Center,
Syracuse, New York 13210
Previous work has revealed a cytoplasmic pool of flagellar
precursor proteins capable of contributing to the assembly of new flagella, but how and where these components assemble is unknown. We
tested Chlamydomonas outer-dynein arm subunit stability
and assembly in the cytoplasm of wild-type cells and 11 outer dynein arm assembly mutant strains (oda1-oda11)
by Western blotting of cytoplasmic extracts, or immunoprecipitates from
these extracts, with five outer-row dynein subunit-specific antibodies.
Western blots reveal that at least three oda mutants
(oda6, oda7, and oda9)
alter the level of a subunit that is not the mutant gene product.
Immunoprecipitation shows that large preassembled flagellar complexes
containing all five tested subunits (three heavy chains and two
intermediate chains) exist within wild-type cytoplasm. When the
preassembly of these subunits was examined in oda
strains, we observed three patterns: complete coassembly
(oda 1, 3, 5, 8, and 10), partial coassembly
(oda7 and oda11), and no coassembly (oda2, 6, and 9) of the four tested subunits with HC
.
Our data, together with previous studies, suggest that flagellar
outer-dynein arms preassemble into a complete Mr 
2 × 106 dynein arm that resides in a cytoplasmic precursor
pool before transport into the flagellar compartment.
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