scholarly journals Neck linker docking is critical for Kinesin-1 force generation in cells but at a cost to motor speed and processivity

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Breane G Budaitis ◽  
Shashank Jariwala ◽  
Dana N Reinemann ◽  
Kristin I Schimert ◽  
Guido Scarabelli ◽  
...  
Keyword(s):  
Optical Trap ◽  
Force Production ◽  
Force Generation ◽  
Motor Speed ◽  
Force Output ◽  
Mechanical Element ◽  
Membrane Bound ◽  
Dynamics Simulations ◽  
Run Lengths ◽  
In Cells

Kinesin force generation involves ATP-induced docking of the neck linker (NL) along the motor core. However, the roles of the proposed steps of NL docking, cover-neck bundle (CNB) and asparagine latch (N-latch) formation, during force generation are unclear. Furthermore, the necessity of NL docking for transport of membrane-bound cargo in cells has not been tested. We generated kinesin-1 motors impaired in CNB and/or N-latch formation based on molecular dynamics simulations. The mutant motors displayed reduced force output and inability to stall in optical trap assays but exhibited increased speeds, run lengths, and landing rates under unloaded conditions. NL docking thus enhances force production but at a cost to speed and processivity. In cells, teams of mutant motors were hindered in their ability to drive transport of Golgi elements (high-load cargo) but not peroxisomes (low-load cargo). These results demonstrate that the NL serves as a mechanical element for kinesin-1 transport under physiological conditions.

scholarly journals Intracellular cargo transport by single-headed kinesin motors

2019 ◽  
Vol 116 (13) ◽  
pp. 6152-6161 ◽  
Author(s):  
Kristin I. Schimert ◽  
Breane G. Budaitis ◽  
Dana N. Reinemann ◽  
Matthew J. Lang ◽  
Kristen J. Verhey
Keyword(s):  
Intracellular Transport ◽  
Motor Coordination ◽  
Optical Trap ◽  
Coiled Coil ◽  
Force Output ◽  
Cellular Assays ◽  
Cellular Events ◽  
Cargo Transport ◽  
In Cells

Kinesin motor proteins that drive intracellular transport share an overall architecture of two motor domain-containing subunits that dimerize through a coiled-coil stalk. Dimerization allows kinesins to be processive motors, taking many steps along the microtubule track before detaching. However, whether dimerization is required for intracellular transport remains unknown. Here, we address this issue using a combination of in vitro and cellular assays to directly compare dimeric motors across the kinesin-1, -2, and -3 families to their minimal monomeric forms. Surprisingly, we find that monomeric motors are able to work in teams to drive peroxisome dispersion in cells. However, peroxisome transport requires minimal force output, and we find that most monomeric motors are unable to disperse the Golgi complex, a high-load cargo. Strikingly, monomeric versions of the kinesin-2 family motors KIF3A and KIF3B are able to drive Golgi dispersion in cells, and teams of monomeric KIF3B motors can generate over 8 pN of force in an optical trap. We find that intracellular transport and force output by monomeric motors, but not dimeric motors, are significantly decreased by the addition of longer and more flexible motor-to-cargo linkers. Together, these results suggest that dimerization of kinesin motors is not required for intracellular transport; however, it enables motor-to-motor coordination and high force generation regardless of motor-to-cargo distance. Dimerization of kinesin motors is thus critical for cellular events that require an ability to generate or withstand high forces.

scholarly journals Author response: Neck linker docking is critical for Kinesin-1 force generation in cells but at a cost to motor speed and processivity

2019 ◽  
Author(s):  
Breane G Budaitis ◽  
Shashank Jariwala ◽  
Dana N Reinemann ◽  
Kristin I Schimert ◽  
Guido Scarabelli ◽  
...  
Keyword(s):  
Force Generation ◽  
Author Response ◽  
Motor Speed ◽  
In Cells

scholarly journals Pathogenic mutations in the kinesin-3 motor KIF1A diminish force generation and movement through allosteric mechanisms

2021 ◽  
Vol 220 (4) ◽  
Author(s):  
Breane G. Budaitis ◽  
Shashank Jariwala ◽  
Lu Rao ◽  
Yang Yue ◽  
David Sept ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Neurodevelopmental Disorders ◽  
Force Generation ◽  
Long Distance ◽  
Long Distance Transport ◽  
Cargo Transport ◽  
Dynamics Simulations ◽  
Run Lengths ◽  
Allosteric Mechanisms ◽  
Distance Transport

The kinesin-3 motor KIF1A functions in neurons, where its fast and superprocessive motility facilitates long-distance transport, but little is known about its force-generating properties. Using optical tweezers, we demonstrate that KIF1A stalls at an opposing load of ~3 pN but more frequently detaches at lower forces. KIF1A rapidly reattaches to the microtubule to resume motion due to its class-specific K-loop, resulting in a unique clustering of force generation events. To test the importance of neck linker docking in KIF1A force generation, we introduced mutations linked to human neurodevelopmental disorders. Molecular dynamics simulations predict that V8M and Y89D mutations impair neck linker docking. Indeed, both mutations dramatically reduce the force generation of KIF1A but not the motor’s ability to rapidly reattach to the microtubule. Although both mutations relieve autoinhibition of the full-length motor, the mutant motors display decreased velocities, run lengths, and landing rates and delayed cargo transport in cells. These results advance our understanding of how mutations in KIF1A can manifest in disease.

scholarly journals Pathogenic Mutations in the Kinesin-3 Motor KIF1A Diminish Force Generation and Movement Through Allosteric Mechanisms

2020 ◽  
Author(s):  
Breane G. Budaitis ◽  
Shashank Jariwala ◽  
Lu Rao ◽  
David Sept ◽  
Kristen J. Verhey ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Motor Domain ◽  
Force Generation ◽  
Long Distance ◽  
C Elegans ◽  
Long Distance Transport ◽  
Dynamics Simulations ◽  
Run Lengths ◽  
Allosteric Mechanisms ◽  
Kinesin Superfamily

ABSTRACTThe kinesin-3 motor KIF1A functions in neurons where its fast and superprocessive motility is thought to be critical for long-distance transport. However, little is known about the force-generating properties of kinesin-3 motors. Using optical tweezers, we demonstrate that KIF1A and its C. elegans homolog UNC-104 undergo force-dependent detachments at ~3 pN and then rapidly reattach to the microtubule to resume motion, resulting in a sawtooth pattern of clustered force generation events that is unique among the kinesin superfamily. Whereas UNC-104 motors stall before detaching, KIF1A motors do not. To examine the mechanism of KIF1A force generation, we introduced mutations linked to human neurodevelopmental disorders, V8M and Y89D, based on their location in structural elements required for force generation in kinesin-1. Molecular dynamics simulations predict that the V8M and Y89D mutations impair docking of the N-terminal (β9) or C-terminal (β10) portions of the neck linker, respectively, to the KIF1A motor domain. Indeed, both mutations dramatically impair force generation of KIF1A but not the motor’s ability to rapidly reattach to the microtubule track. Homodimeric and heterodimeric mutant motors also display decreased velocities, run lengths, and landing rates and homodimeric Y89D motors exhibit a higher frequency of non-productive, diffusive events along the microtubule. In cells, cargo transport by the mutant motors is delayed. Our work demonstrates the importance of the neck linker in the force generation of kinesin-3 motors and advances our understanding of how mutations in the kinesin motor domain can manifest in disease.

scholarly journals Neurophysiological changes in the visuomotor network after practicing a motor task

2018 ◽  
Vol 120 (1) ◽  
pp. 239-249 ◽  
Author(s):  
James E. Gehringer ◽  
David J. Arpin ◽  
Elizabeth Heinrichs-Graham ◽  
Tony W. Wilson ◽  
Max J. Kurz
Keyword(s):  
Motor Learning ◽  
Motor Planning ◽  
Motor Task ◽  
Force Production ◽  
Oscillatory Activity ◽  
Matching Task ◽  
Force Output ◽  
Motor Action ◽  
Visuomotor Transformations ◽  
Target Matching

Although it is well appreciated that practicing a motor task updates the associated internal model, it is still unknown how the cortical oscillations linked with the motor action change with practice. The present study investigates the short-term changes (e.g., fast motor learning) in the α- and β-event-related desynchronizations (ERD) associated with the production of a motor action. To this end, we used magnetoencephalography to identify changes in the α- and β-ERD in healthy adults after participants practiced a novel isometric ankle plantarflexion target-matching task. After practicing, the participants matched the targets faster and had improved accuracy, faster force production, and a reduced amount of variability in the force output when trying to match the target. Parallel with the behavioral results, the strength of the β-ERD across the motor-planning and execution stages was reduced after practice in the sensorimotor and occipital cortexes. No pre/postpractice changes were found in the α-ERD during motor planning or execution. Together, these outcomes suggest that fast motor learning is associated with a decrease in β-ERD power. The decreased strength likely reflects a more refined motor plan, a reduction in neural resources needed to perform the task, and/or an enhancement of the processes that are involved in the visuomotor transformations that occur before the onset of the motor action. These results may augment the development of neurologically based practice strategies and/or lead to new practice strategies that increase motor learning. NEW & NOTEWORTHY We aimed to determine the effects of practice on the movement-related cortical oscillatory activity. Following practice, we found that the performance of the ankle plantarflexion target-matching task improved and the power of the β-oscillations decreased in the sensorimotor and occipital cortexes. These novel findings capture the β-oscillatory activity changes in the sensorimotor and occipital cortexes that are coupled with behavioral changes to demonstrate the effects of motor learning.

Intermittency in the Control of Continuous Force Production

2000 ◽  
Vol 84 (4) ◽  
pp. 1708-1718 ◽  
Author(s):  
Andrew B. Slifkin ◽  
David E. Vaillancourt ◽  
Karl M. Newell
Keyword(s):  
Visual Feedback ◽  
Visual Information ◽  
Spectral Power ◽  
Force Production ◽  
Motor Output ◽  
Force Variability ◽  
Force Output ◽  
Feedback Information ◽  
Information Processes ◽  
Feedback Frequency

The purpose of the current investigation was to examine the influence of intermittency in visual information processes on intermittency in the control continuous force production. Adult human participants were required to maintain force at, and minimize variability around, a force target over an extended duration (15 s), while the intermittency of on-line visual feedback presentation was varied across conditions. This was accomplished by varying the frequency of successive force-feedback deliveries presented on a video display. As a function of a 128-fold increase in feedback frequency (0.2 to 25.6 Hz), performance quality improved according to hyperbolic functions (e.g., force variability decayed), reaching asymptotic values near the 6.4-Hz feedback frequency level. Thus, the briefest interval over which visual information could be integrated and used to correct errors in motor output was approximately 150 ms. The observed reductions in force variability were correlated with parallel declines in spectral power at about 1 Hz in the frequency profile of force output. In contrast, power at higher frequencies in the force output spectrum were uncorrelated with increases in feedback frequency. Thus, there was a considerable lag between the generation of motor output corrections (1 Hz) and the processing of visual feedback information (6.4 Hz). To reconcile these differences in visual and motor processing times, we proposed a model where error information is accumulated by visual information processes at a maximum frequency of 6.4 per second, and the motor system generates a correction on the basis of the accumulated information at the end of each 1-s interval.

Neuromuscular drive and force production are not altered during bilateral contractions

1998 ◽  
Vol 84 (1) ◽  
pp. 200-206 ◽  
Author(s):  
J. M. Jakobi ◽  
E. Cafarelli
Keyword(s):  
Motor Unit ◽  
Force Production ◽  
Young Male ◽  
Neuromuscular Control ◽  
Force Generation ◽  
Isometric Contractions ◽  
Firing Rates ◽  
Significant Limitation ◽  
Motor Unit Firing ◽  
Male Subjects

Jakobi, J. M., and E. Cafarelli. Neuromuscular drive and force production are not altered during bilateral contractions. J. Appl. Physiol. 84(1): 200–206, 1998.—Several investigators have studied the deficit in maximal voluntary force that is said to occur when bilateral muscle groups contract simultaneously. A true bilateral deficit (BLD) would suggest a significant limitation of neuromuscular control; however, some of the data from studies in the literature are equivocal. Our purpose was to determine whether there is a BLD in the knee extensors of untrained young male subjects during isometric contractions and whether this deficit is associated with a decreased activation of the quadriceps, increased activation of the antagonist muscle, or an alteration in motor unit firing rates. Twenty subjects performed unilateral (UL) and bilateral (BL) isometric knee extensions at 25, 50, 75, and 100% maximal voluntary contraction. Total UL and BL force (Δ3%) and maximal rate of force generation (Δ2.5%) were not significantly different. Total UL and BL maximal vastus lateralis electromyographic activity (EMG; 2.7 ± 0.28 vs. 2.6 ± 0.24 mV) and coactivation (0.17 ± 0.02 vs. 0.20 ± 0.02 mV) were also not different. Similarly, the ratio of force to EMG during submaximal UL and BL contractions was not different. Analysis of force production by each leg in UL and BL conditions showed no differences in force, rate of force generation, EMG, motor unit firing rates, and coactivation. Finally, assessment of quadriceps activity with the twitch interpolation technique indicated no differences in the degree of voluntary muscle activation (UL: 93.6 ± 2.51 Hz, BL: 90.1 ± 2.43 Hz). These results provide no evidence of a significant limitation in neuromuscular control between BL and UL isometric contractions of the knee extensor muscles in young male subjects.

Effects of hypoxia and hypercapnia on geniohyoid contractility and endurance

1991 ◽  
Vol 71 (2) ◽  
pp. 709-715 ◽  
Author(s):  
R. J. Salmone ◽  
E. Van Lunteren
Keyword(s):  
Force Production ◽  
Upper Airway ◽  
Muscle Performance ◽  
Severe Hypoxia ◽  
Force Frequency ◽  
Force Output ◽  
Frequency Curve ◽  
Dilator Muscle ◽  
Index Ratio ◽  
Mild Hypoxia

Sleep apnea and other respiratory diseases produce hypoxemia and hypercapnia, factors that adversely affect skeletal muscle performance. To examine the effects of these chemical alterations on force production by an upper airway dilator muscle, the contractile and endurance characteristics of the geniohyoid muscle were examined in situ during severe hypoxia (arterial PO2 less than 40 Torr), mild hypoxia (PO2 45–65 Torr), and hypercapnia (PCO2 55–80 Torr) and compared with hyperoxic-normocapnic conditions in anesthetized cats. Muscles were studied at optimal length, and contractile force was assessed in response to supramaximal electrical stimulation of the hypoglossal nerve (n = 7 cats) or geniohyoid muscle (n = 2 cats). There were no significant changes in the twitch kinetics or force-frequency curve of the geniohyoid muscle during hypoxia or hypercapnia. However, the endurance of the geniohyoid, as reflected in the fatigue index (ratio of force at 2 min to initial force in response to 40-Hz stimulation at a duty cycle 0.33), was significantly reduced by severe hypoxia but not by hypercapnia or mild hypoxia. In addition, the downward shift in the force-frequency curve after the repetitive stimulation protocol was greater during hypoxia than hyperoxia, especially at higher frequencies. In conclusion, the ability of the geniohyoid muscle to maintain force output during high levels of activation is adversely affected by severe hypoxia but not mild hypoxia or hypercapnia. However, none of these chemical perturbations affected muscle contractility acutely.

scholarly journals Force Generation by Membrane-Bound Myo1c, a Single Molecule Study

2013 ◽  
Vol 104 (2) ◽  
pp. 642a
Author(s):  
Serapion Pyrpassopoulos ◽  
Henry Shuman ◽  
E. Michael Ostap
Keyword(s):  
Single Molecule ◽  
Force Generation ◽  
Membrane Bound
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