Abstract
BackgroundKinesin 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.ResultsWe 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.ConclusionThe 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.
Highlights
Kinesin and dynein are the two families of microtubule-based motors that drive much of the intracellular movements in eukaryotic cells
Characterization of the actin cytoskeleton in Dictyostelium has led to the identification of actin binding proteins, multiple myosin motors, and signaling cascades whose functions are conserved among eukaryotic cells
We address here the consequences of individual disruption of four kinesin genes in Dictyostelium, and we contrast the functional redundancies among such motors in single-celled organisms with their functional specificity in metazoan organisms
Summary
Kinesin and dynein are the two families of microtubule-based motors that drive much of the intracellular movements in eukaryotic cells. When food sources are exhausted, individual amoebae trigger a developmental program that initiates both inter and intracellular signaling, to aggregate ~100,000 amoebae and form a multicellular mass. Each cell within this mass undergoes multiple adhesions and conformational changes, forming a cooperative slug that can migrate to new areas. Motility-wise, Dictyostelium behaves in a manner similar to that of many vertebrate cells (crawling, sensing, and engulfing targets, robust intracellular movements) This organism clearly retains a simplicity associated with its relatively small and compact genome, and exhibits features commonly seen in protozoa and fungi (for example, an intranuclear spindle for cell division). The machinery in Dictyostelium that drives movement along microtubules contains 14 motors
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