Disruption of genes encoding predicted inner arm dynein heavy chains causes motility phenotypes in Tetrahymena.
The multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132] states: (1) there are many different dynein HC isoforms; (2) each isoform is encoded by a different gene; (3) different isoforms have different functions. Many studies provide evidence in support of the first two statements [Piperno et al., 1990: J Cell Biol 110:379-389; Kagami and Kamiya, 1992: J Cell Sci 103:653-664; Gibbons, 1995: Cell Motil Cytoskeleton 32:136-144; Porter et al., 1996: Genetics 144:569-585; Xu et al., 1999: J Eukaryot Microbiol 46:606-611] and there is evidence that outer arms and inner arms play different roles in flagellar beating [Brokaw and Kamiya, 1987: Cell Motil. Cytoskeleton 8:68-75]. However, there are few studies rigorously testing in vivo whether inner arm dyneins, especially the 1-headed inner arm dyneins, play unique roles. This study tested the third tenet of the multi-dynein hypothesis by introducing mutations into three inner arm dynein HC genes (DYH8, 9 and 12) that are thought to encode HCs associated with 1-headed inner arm dyneins. Southern blots, Northern blots, and RT-PCR analyses indicate that all three mutants (KO-8, 9, and 12) are complete knockouts. Each mutant swims slower than the wild-type cells. The beat frequency of KO-8 cells is lower than that of the wild-type cells while the beat frequencies of KO-9 and KO-12 are not different from that of wild-type cells. Our results suggest that each inner arm dynein HC is essential for normal cell motility and cannot be replaced functionally by other dynein HCs and that not all of the 1-headed inner arm dyneins play the same role in ciliary motility. Thus, the results of our study support the multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132].
Department of Zoology, Miami University, Oxford, OH 45056, USA.
Siming
S
Liu
Robert
R
Hard
Scott
S
Rankin
Todd
T
Hennessey
David G
DG
Pennock
D000818:Animals; D030801:Animals, Genetically Modified; D002465:Cell Movement / Q000502:physiology*; D002923:Cilia / Q000378:metabolism; D004398:Dyneins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018390:Gene Targeting; D010641:Phenotype; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016808:Tetrahymena thermophila / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*
D020033:Protein Isoforms; D015800:Protozoan Proteins; D004398:Dyneins