An extended microtubule-binding structure within the dynein motor domain

Gee, Melissa A.; Heuser, John E.; Vallee, Richard B.

doi:10.1038/37663

Cited by 274 publications

(278 citation statements)

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“…Analysis of dynein sequences from various organisms previously predicted the stalk to be composed of an antiparallel coiled-coil extending to the two strictly conserved prolines at either end of the MTDB headpiece (6,7,10,42). However, as revealed by x-ray analysis of the murine MTBD (1), the distal end of the coiled-coil extends past these prolines to form one face of the MTBD itself.…”

Section: Backbone Chemical Shift Analysis Of the Isolated D Discoidementioning

confidence: 99%

“…For dynein, these issues are particularly complex. In contrast to the close proximity of the substrate-binding domains and catalytic sites in kinesin or myosin-type motors (5), the ATP-sensitive interaction of dynein with microtubules occurs through contacts within a relatively small (ϳ125 residue) globular domain (microtubule binding domain, MTBD), 2 located at the distal end of a ϳ10-nm long ␣-helical antiparallel coiled-coil stalk (6,7). This stalk projects off the roughly 3000-residue ring-shaped AAAϩ motor domain (Fig.…”

mentioning

confidence: 99%

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A Low Affinity Ground State Conformation for the Dynein Microtubule Binding Domain

McNaughton

Tikhonenko

Banavali

et al. 2010

Journal of Biological Chemistry

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Dynein interacts with microtubules through a dedicated binding domain that is dynamically controlled to achieve high or low affinity, depending on the state of nucleotide bound in a distant catalytic pocket. The active sites for microtubule binding and ATP hydrolysis communicate via conformational changes transduced through a ϳ10-nm length antiparallel coiled-coil stalk, which connects the binding domain to the roughly 300-kDa motor core. Recently, an x-ray structure of the murine cytoplasmic dynein microtubule binding domain (MTBD) in a weak affinity conformation was published, containing a covalently constrained ␤ ؉ registry for the coiled-coil stalk segment (Carter, A. P., Garbarino, J. E., Wilson-Kubalek, E. M., Shipley, W. E., Cho, C., Milligan, R. A., Vale, R. D., and Gibbons, I. R. (2008) Science 322, 1691-1695). We here present an NMR analysis of the isolated MTBD from Dictyostelium discoideum that demonstrates the coiled-coil ␤ ؉ registry corresponds to the low energy conformation for this functional region of dynein. Addition of sequence encoding roughly half of the coiled-coil stalk proximal to the binding tip results in a decreased affinity of the MTBD for microtubules. In contrast, addition of the complete coiled-coil sequence drives the MTBD to the conformationally unstable, high affinity binding state. These results suggest a thermodynamic coupling between conformational free energy differences in the ␣ and ␤ ؉ registries of the coiled-coil stalk that acts as a switch between high and low affinity conformations of the MTBD. A balancing of opposing conformations in the stalk and MTBD enables potentially modest long-range interactions arising from ATP binding in the motor core to induce a relaxation of the MTBD into the stable low affinity state.Dyneins comprise one of the three families of cytoskeletonbased molecular motors that generate force and translocate cargo in eukaryotic cells (2). These motors work by coupling high and low affinity binding to microtubule or actin filaments, with force producing conformational changes driven by an ATP catalytic cycle (3-5). The coordination between these steps is critical for efficient linear movement. Force production occurs after tight binding is achieved, in a manner that both moves cargo forward and facilitates motor repositioning to advance another step. Although ATP hydrolysis and product release provide the thermodynamic driving force for motility, binding to the microtubule or actin filaments also provides feedback to the catalytic pocket and influences the catalytic rate.An understanding of how substrate affinity is achieved and how it is coupled to nucleotide hydrolysis remains an important problem for all three families of motors (dynein, kinesin, and myosin). For dynein, these issues are particularly complex. In contrast to the close proximity of the substrate-binding domains and catalytic sites in kinesin or myosin-type motors (5), the ATP-sensitive interaction of dynein with microtubules occurs through contacts within a relatively small (ϳ125 ...

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Section: Backbone Chemical Shift Analysis Of the Isolated D Discoidementioning

confidence: 99%

mentioning

confidence: 99%

A Low Affinity Ground State Conformation for the Dynein Microtubule Binding Domain

McNaughton

Tikhonenko

Banavali

et al. 2010

Journal of Biological Chemistry

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“…These two structures bind the microtubule track and cargo, respectively 7,12,13 . The stalk is up to 15 nm in length in some species 10 and is most probably an anti-parallel coiled-coil 14 . A small globular domain at its tip is the ATP-sensitive microtubule-binding domain [14][15][16] .…”

mentioning

confidence: 99%

“…The stalk is up to 15 nm in length in some species 10 and is most probably an anti-parallel coiled-coil 14 . A small globular domain at its tip is the ATP-sensitive microtubule-binding domain [14][15][16] . The stalk and head are formed by the carboxyterminal two-thirds of the heavy chain 14,17 .…”

mentioning

confidence: 99%

“…A small globular domain at its tip is the ATP-sensitive microtubule-binding domain [14][15][16] . The stalk and head are formed by the carboxyterminal two-thirds of the heavy chain 14,17 . The stem, which also binds the intermediate and light chains 7 , is formed by the aminoterminal region and contains numerous short stretches with predicted coiled-coil structure 18,19 .…”

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Dynein structure and power stroke

Burgess

Walker

Sakakibara

et al. 2003

Nature

478

552

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Dynein ATPases are microtubule motors that are critical to diverse processes such as vesicle transport and the beating of sperm tails; however, their mechanism of force generation is unknown. Each dynein comprises a head, from which a stalk and a stem emerge. Here we use electron microscopy and image processing to reveal new structural details of dynein c, an isoform from Chlamydomonas reinhardtii flagella, at the start and end of its power stroke. Both stem and stalk are flexible, and the stem connects to the head by means of a linker approximately 10 nm long that we propose lies across the head. With both ADP and vanadate bound, the stem and stalk emerge from the head 10 nm apart. However, without nucleotide they emerge much closer together owing to a change in linker orientation, and the coiled-coil stalk becomes stiffer. The net result is a shortening of the molecule coupled to an approximately 15-nm displacement of the tip of the stalk. These changes indicate a mechanism for the dynein power stroke.

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Dynein and dynein-related genes

Milisav

1998

Cell Motil. Cytoskeleton

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An extended microtubule-binding structure within the dynein motor domain

Cited by 274 publications

References 29 publications

A Low Affinity Ground State Conformation for the Dynein Microtubule Binding Domain

A Low Affinity Ground State Conformation for the Dynein Microtubule Binding Domain

Dynein structure and power stroke

Dynein and dynein-related genes

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