Disruption of four kinesin genes in dictyostelium.
BACKGROUND
Kinesin and dynein are the two families of microtubule-based motors that drive much of the intracellular movements in eukaryotic cells. Using a gene knockout strategy, we address here the individual function(s) of four of the 13 kinesin proteins in Dictyostelium. The goal of our ongoing project is to establish a minimal motility proteome for this basal eukaryote, enabling us to contrast motor functions here with the often far more elaborate motor families in the metazoans.


RESULTS
We performed individual disruptions of the kinesin genes, kif4, kif8, kif10, and kif11. None of the motors encoded by these genes are essential for development or viability of Dictyostelium. Removal of Kif4 (kinesin-7; CENP-E family) significantly impairs the rate of cell growth and, when combined with a previously characterized dynein inhibition, results in dramatic defects in mitotic spindle assembly. Kif8 (kinesin-4; chromokinesin family) and Kif10 (kinesin-8; Kip3 family) appear to cooperate with dynein to organize the interphase radial microtubule array.


CONCLUSIONS
The results reported here extend the number of kinesin gene disruptions in Dictyostelium, to now total 10, among the 13 isoforms. None of these motors, individually, are required for short-term viability. In contrast, homologs of at least six of the 10 kinesins are considered essential in humans. Our work underscores the functional redundancy of motor isoforms in basal organisms while highlighting motor specificity in more complex metazoans. Since motor disruption in Dictyostelium can readily be combined with other motility insults and stresses, this organism offers an excellent system to investigate functional interactions among the kinesin motor family.
Nag
Dilip K
DK
Division of Molecular Medicine, Wadsworth Center, Albany, NY 12201-0509, USA. nagd@wadsworth.org
Tikhonenko
Irina
I
Soga
Ikko
I
Koonce
Michael P
MP
D000818:Animals; D004023:Dictyostelium / Q000201:enzymology / Q000235:genetics*; D004398:Dyneins / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D016547:Kinesins / Q000235:genetics* / Q000378:metabolism; D008870:Microtubules / Q000201:enzymology / Q000648:ultrastructure; D010802:Phylogeny; D017384:Sequence Deletion; D008941:Spindle Apparatus / Q000201:enzymology / Q000648:ultrastructure; D019076:Transgenes
D005656:Fungal Proteins; D004398:Dyneins; D016547:Kinesins