Targeted gene disruption of dynein heavy chain 7 of Tetrahymena thermophila results in altered ciliary waveform and reduced swim speed.
Tetrahymena thermophila swims by the coordinated beating of hundreds of cilia that cover its body. It has been proposed that the outer arm dyneins of the ciliary axoneme control beat frequency, whereas the inner arm dyneins control waveform. To test the role of one of these inner arms, dynein heavy chain 7 protein (Dyh7p), a knockout mutant was generated by targeted biolistic transformation of the vegetative macronucleus. Disruption of DYH7, the gene which encodes Dyh7p, was confirmed by PCR examination of both genomic and cDNA templates. Both intact and detergent extracted, reactivated cell model preparations of these mutants, which we call DYH7neo3, displayed swim speeds that were almost half that of wild-type cells. Although the DYH7neo3 mutants were slower than wild type, they were able to modulate their swim speed and show ciliary reversal in response to depolarizing stimuli. High-speed video microscopy of intact, free-swimming DYH7neo3 mutants revealed an irregular pattern of ciliary beat and waveform. The mutant cilia appeared to be engaging in less coordinated, swiveling movements in which the typical shape, periodicity and coordination seen in wild-type cilia were absent or disturbed. We propose that the axonemal inner arm dynein heavy chain 7 proteins contribute to the formation of normal ciliary waveform, which in turn governs the forward swimming velocity of these cells.
Department of Biological Sciences, State University of New York at Buffalo, Amherst, NY 14260, USA.
Christopher R
CR
Wood
Robert
R
Hard
Todd M
TM
Hennessey
D000818:Animals; D001522:Behavior, Animal / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002923:Cilia / Q000378:metabolism* / Q000648:ultrastructure; D003902:Detergents / Q000737:chemistry; D004398:Dyneins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D018390:Gene Targeting; D005838:Genotype; D010641:Phenotype; D010802:Phylogeny; D015800:Protozoan Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D013550:Swimming; D016808:Tetrahymena thermophila / Q000502:physiology*
D003902:Detergents; D015800:Protozoan Proteins; D004398:Dyneins