The Directional Preference of Kinesin Motors Is Specified by an Element outside of the Motor Catalytic Domain

Case, Ryan; Pierce, Daniel W.; Hom-Booher, Nora; Hart, Cynthia L.; Vale, Ronald D.

doi:10.1016/s0092-8674(00)80360-8

Cited by 343 publications

(333 citation statements)

References 42 publications

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“…E311 (in the R6 helix) immediately precedes the neck linker (4), a region which has been implicated in mechanical amplification and directional motion (27,31,43). We speculate that motions of R6 could affect the position and conformation of the neck linker.…”

Section: Discussionmentioning

confidence: 98%

“…In a multiple motor assay, several motors interact with a microtubule while coupled to a common mechanical substrate, the slide. This enables them to work cooperatively, and as a result, many nonprocessive motors show robust movement in multiple motor assays, while showing no motility in single molecule assays (27,45). In the single motor assay, there is the additional requirement that the two heads of the dimer coordinate their actions to ensure that one head is attached to the microtubule at all times.…”

Section: Discussionmentioning

confidence: 99%

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ATPase Kinetic Characterization and Single Molecule Behavior of Mutant Human Kinesin Motors Defective in Microtubule-Based Motility

et al. 2000

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Conventional kinesin is a microtubule-based motor protein that is an important model system for understanding mechanochemical transduction. To identify regions of the kinesin protein that participate in microtubule binding and force production, Woehlke et al. [(1997) Cell 90, 207-216] generated 35 alanine mutations in solvent-exposed residues. Here, we have performed presteady-state kinetic and single molecule motility analyses on three of these mutants [Y138A, loop 11 triple (L248A/D249A/E250A), and E311A] that exhibited a similar ∼3-fold reduction in both microtubule gliding velocity and microtubulestimulated ATPase activity. All mutants showed normal second-order ATP binding kinetics, indicating correct folding of the active site. The Y138A and loop 11 triple mutants were defective both in nucleotide hydrolysis and in microtubule-stimulated ADP release rates, the latter suggesting a defect in allosteric communication between the microtubule and the active site. A single molecule fluorescence assay further revealed that the loop 11 mutant is defective in initiating processive motion, suggesting that this loop is important for the initial contact between kinesin and the microtubule. Y138A, on the other hand, can bind to the microtubule normally but cannot move processively. For E311A, neither the rate of nucleotide hydrolysis nor ADP release could account for its slower ATPase and gliding velocity, which suggests that either phosphate release or a conformational transition is rate-limiting in this mutant. The single molecule assay showed that E311A has a reduced velocity of movement, but is not defective in processivity. Thus, while these mutants behave similarly in solution ATPase and multiple motor gliding assays, kinetic and single molecule analyses reveal defects in distinct processes in kinesin's mechanochemical cycle.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

ATPase Kinetic Characterization and Single Molecule Behavior of Mutant Human Kinesin Motors Defective in Microtubule-Based Motility

et al. 2000

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“…24 Recombinant conventional kinesin-1 consisting of first 573 amino acid residues of human kinesin-1 (K573) and GFP-fused kinesin-1 construct consisting of the first 560 amino acid residues of human kinesin-1 (K560) were prepared as described in previously published papers by partially modifying the expression and purification methods. 25 …”

Section: Experimental Section Preparation Of Tubulin and Kinesinmentioning

confidence: 99%

Effect of length and rigidity of microtubules on the size of ring-shaped assemblies obtained through active self-organization

et al. 2015

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The microtubule (MT)-kinesin biomolecular motor system has attracted much attention due to its possible applications in artificial biomachines. Recently an active self-organization (AcSO) method has been established to integrate MT filaments into highly organized assembled structures. The ring-shaped MT assembly, one of the structures derived from the AcSO of MTs, can convert the translational motion of MTs into rotational motion. Due to this attractive feature, the ring-shaped MT assembly appears to be a promising candidate for developing artificial devices and for future nanotechnological applications. In this work, we have investigated the effect of length and rigidity of MT filaments on the size of ring-shaped MT assembly in the AcSO process. We show that the size of ring-shaped MT assembly can be controlled by tuning the length and rigidity of MT filaments employed in the AcSO.Longer and stiffer MT filaments led to larger ring-shaped assemblies through AcSO, while AcSO of shorter and less stiff MT filaments produced smaller ring-shaped assemblies. This work might be important for the development of biomolecular motor based artificial biomachines, especially where size control of ring-shaped MT assembly will play an important role.

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“…The R350 amino acid in particular is located at the end of the motor domain in close vicinity to the neck linker (aa 352-364) that has an important role in directionality and has mechanochemical implications for the motility of the protein. [7][8][9] There is evidence that the KIF1A protein functions as the primary motor for synaptic-and dense-core vesicle transport. 10,11 In animal models of C. elegans with mutations in the KIF1A ortholog (unc-104), a decreased transport capacity of synaptic vesicle precursors in the axons has been evidenced.…”

Section: Rearrangement Detectionmentioning

confidence: 99%

KIF1A missense mutations in SPG30, an autosomal recessive spastic paraplegia: distinct phenotypes according to the nature of the mutations

et al. 2012

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The hereditary spastic paraplegias (HSPs) are a clinically and genetically heterogeneous group of neurodegenerative diseases characterised by progressive spasticity in the lower limbs. The nosology of autosomal recessive forms is complex as most mapped loci have been identified in only one or a few families and account for only a small percentage of patients. We used next-generation sequencing focused on the SPG30 chromosomal region on chromosome 2q37.3 in two patients from the original linked family. In addition, wide genome scan and candidate gene analysis were performed in a second family of Palestinian origin. We identified a single homozygous mutation, p.R350G, that was found to cosegregate with the disease in the SPG30 kindred and was absent in 970 control chromosomes while affecting a strongly conserved amino acid at the end of the motor domain of KIF1A. Homozygosity and linkage mapping followed by mutation screening of KIF1A allowed us to identify a second mutation, p.A255V, in the second family. Comparison of the clinical features with the nature of the mutations of all reported KIF1A families, including those reported recently with hereditary sensory and autonomic neuropathy, suggests phenotypegenotype correlations that may help to understand the mechanisms involved in motor neuron degeneration. We have shown that mutations in the KIF1A gene are responsible for SPG30 in two autosomal recessive HSP families. In published families, the nature of the KIF1A mutations seems to be of good predictor of the underlying phenotype and vice versa. INTRODUCTIONThe hereditary spastic paraplegias (HSPs) are a clinically and genetically heterogeneous group of neurodegenerative diseases characterised by progressive spasticity in the lower limbs. 1 The mode of inheritance may be autosomal dominant, autosomal recessive (ARHSP) or X-linked. More than 48 different loci (SPGn) have been mapped so far, and 23 responsible genes identified. The corresponding proteins are often involved in intracellular trafficking or mitochondrial functions. 2,3 Clinically, one can distinguish between pure and complicated forms of HSP. 1,2 Pure forms consist of isolated pyramidal signs, such as spasticity, abnormal reflexes (brisk reflexes and Babinski sign) and motor deficit, often associated with sphincter disturbances and deep sensory loss. In the complicated forms of HSP the disease is variably associated with numerous combinations of neurological and extraneurological signs, such as cerebellar ataxia, dysarthria, mental retardation, peripheral neuropathy, optic atrophy, retinitis pigmentosa and/or hearing loss, which may be accompanied by abnormal brain MRI (atrophy of the cortex, cerebellum or corpus callosum, white matter abnormalities, etc).Mutations in the KIF1A gene (MIM 601255) were very recently described in two different clinically and genetically heterogeneous groups of neurodegenerative diseases. 4,5 In hereditary sensory and autonomic neuropathy (HSAN), all patients from four families with different origins shared a common hom...

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The Directional Preference of Kinesin Motors Is Specified by an Element outside of the Motor Catalytic Domain

Cited by 343 publications

References 42 publications

ATPase Kinetic Characterization and Single Molecule Behavior of Mutant Human Kinesin Motors Defective in Microtubule-Based Motility

ATPase Kinetic Characterization and Single Molecule Behavior of Mutant Human Kinesin Motors Defective in Microtubule-Based Motility

Effect of length and rigidity of microtubules on the size of ring-shaped assemblies obtained through active self-organization

KIF1A missense mutations in SPG30, an autosomal recessive spastic paraplegia: distinct phenotypes according to the nature of the mutations

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