The Effect of Monastrol on the Processive Motility of a Dimeric Kinesin-5 Head/Kinesin-1 Stalk Chimera

Lakämper, Stefan; Thiede, Christina; Düselder, André; Reiter, Stefanie; Korneev, M.; Kapitein, Lukas C.; Peterman, Erwin J. G.; Schmidt, Christoph F.

doi:10.1016/j.jmb.2010.03.009

Cited by 25 publications

(40 citation statements)

References 34 publications

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“…3C), whereas the speed remained slow at 0.08 AE 0.02 μm∕s (mean AE SD of mean). This result is similar to results from Lakamper et al (31) who studied a similar construct. This result demonstrates that, to the degree that there are any unique kinetic characteristics of kinesin-5 that result in low processivity in the wild-type motor, they can be completely compensated for by shortening the neck linker to enhance head-head coordination.…”

Section: Resultssupporting

confidence: 92%

Interhead tension determines processivity across diverse N-terminal kinesins

Shastry

Hancock

2011

Proc. Natl. Acad. Sci. U.S.A.

134

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Consistent with their diverse intracellular roles, the processivity of N-terminal kinesin motors varies considerably between different families. Kinetics experiments on isolated motor domains suggest that differences in processivity result from differences in the underlying biochemistry of the catalytic heads. However, the length of the flexible neck linker domain also varies from 14 to 18 residues between families. Because the neck linker acts as a mechanical element that transmits interhead tension, altering its mechanical properties is expected to affect both front and rear head gating, mechanisms that underlie processive walking. To test the hypothesis that processivity differences result from family-specific differences in neck linker mechanics, we systematically altered the neck linker length in kinesin-1, -2, -3, -5, and -7 motors and measured run length and velocity in a single-molecule fluorescence assay. Shortening the neck linkers of kinesin-3 (Unc104/KIF1A) and kinesin-5 (Eg5/KSP) to 14 residues enhanced processivity to match kinesin-1, which has a 14-residue neck linker. After substituting a single residue in the last alpha helix of the catalytic core, kinesin-7 (CENP-E) exhibited this same behavior. This convergence of processivity was observed even though motor speeds varied over a 25-fold range. These results suggest that differences in unloaded processivity between diverse kinesins is primarily due to differences in the lengths of their neck linker domains rather than specific tuning of rate constants in their ATP hydrolysis cycles.cytoskeleton | molecular motor | intracellular transport | Monte Carlo model

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Section: Resultssupporting

confidence: 92%

Interhead tension determines processivity across diverse N-terminal kinesins

Shastry

Hancock

2011

Proc. Natl. Acad. Sci. U.S.A.

134

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“…1A). For Eg5 kinesin-5 (14,15), this strategy had provided a stable dimeric chimera capable of processive motility.…”

Section: Resultsmentioning

confidence: 99%

“…Plus-end-directed motility was demonstrated in vitro for a number of kinesin-5 motors in MT surface gliding assays (8), in single-molecule fluorescence motility assays (14,15), and in MT sliding assays (9,16). Surprisingly, single molecules of Saccharomyces cerevisiae kinesin-5 Cin8 were recently shown to move in the minus-end direction of the MTs, in highionic-strength conditions (17)(18)(19).…”

mentioning

confidence: 99%

Deletion of the Tail Domain of the Kinesin-5 Cin8 Affects Its Directionality

Düselder

Fridman²,

Thiede

et al. 2015

Journal of Biological Chemistry

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Background: Single molecules of the kinesin-5 Cin8 were previously demonstrated to be minus-end-directed under highionic-strength conditions. Results: Under high-ionic-strength conditions, Cin8 lacking the tail domain is bidirectional. Conclusion:The tail domain is one of the factors that regulate Cin8 directionality. Significance: An important structural element was identified that regulates the directionality of kinesin-5 motors.

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“…34 The initial rationale for using 14 amino acids was to provide a similar mechanical background as in Kinesin-1. We had found that Eg5Kin (here referred to as Eg5Kin-NL-14 or, short, NL-14) is capable of long-range processive motility taking on average 240 consecutive~8-nm steps and of comparatively high force generation.…”

Section: Introductionmentioning

confidence: 99%