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Vol. 9, Issue 12, 3351-3365, December 1998
and
*Department of Cell Biology and Neuroanatomy, University of
Minnesota Medical School, Minneapolis, Minnesota 55455;
To identify new loci that are involved in the assembly and
targeting of dynein complexes, we have screened a collection of motility mutants that were generated by insertional mutagenesis. One
such mutant, 5B10, lacks the inner arm isoform known as the I1 complex.
This isoform is located proximal to the first radial spoke in each
96-nm axoneme repeat and is an important target for the regulation of
flagellar motility. Complementation tests reveal that 5B10 represents a
new I1 locus, IDA7. Biochemical analyses confirm that
ida7 axonemes lack at least five I1 complex subunits.
Southern blots probed with a clone containing the gene encoding the
140-kDa intermediate chain (IC) indicate that the ida7 mutation is the result of plasmid insertion into
the IC140 gene. Transformation with a wild-type copy of the IC140 gene
completely rescues the mutant defects. Surprisingly, transformation
with a construct of the IC140 gene lacking the first four exons of the
coding sequence also rescues the mutant phenotype. These studies indicate that IC140 is essential for assembly of the I1 complex, but
unlike other dynein ICs, the N-terminal region is not critical for its activity.
Department of Anatomy and Cell Biology, Emory University
Medical School, Atlanta, Georgia 30322; and
Department
of Molecular, Cellular, and Developmental Biology, University of
Colorado at Boulder, Boulder, Colorado 80309-0347
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