Structural basis for the extended CAP-Gly domains of p150
            <sup>glued</sup>
            binding to microtubules and the implication for tubulin dynamics

Wang, Qianmin; Crevenna, Álvaro H.; Kunze, I; Mizuno, Naoko

doi:10.1073/pnas.1403135111

Cited by 43 publications

(67 citation statements)

References 34 publications

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“…Our current results also clearly indicate that the bean-shaped CAP-Gly molecule binds to the microtubules with its flat surface comprised by its loops, the C-terminal β4-strand, and the short α-helix. These findings are in agreement with the recent cryo-EM results on the CAP-Gly(1-105)/MT and CAP-Gly(25-144)/MT complexes (40), which revealed the same flat side of CAP-Gly that forms the interface, including the proposed interaction of the GKNDG motif with the C terminus of tubulin. Of note, most of the surface-exposed, hydrophobic residues are on the flat side of CAP-Gly and are likely important for promoting the binding interaction.…”

Section: Resultssupporting

confidence: 92%

“…Recently, a cryo-EM study reported the structure of several CAP-Gly constructs on polymerized MTs at 9.7-10.2 Å resolution (40). Although atomic resolution was not achieved in this study, several important observations were made: (i) The interface of CAP-Gly with MT includes the GKNDG motif, which interacts with the C terminus of tubulin; (ii) the tubulin E-hooks interact with the basic patches 1-25 and 106-144 of the protein, possibly permitting it to diffuse laterally to the next binding site; (iii) the orientation of CAP-Gly on MTs depends on the length of the construct used; and (iv) the recognition sites for CAP-Gly and EB1 on tubulin are nonoverlapping.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Yan

Guo

Hou

et al. 2015

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

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Microtubules and their associated proteins perform a broad array of essential physiological functions, including mitosis, polarization and differentiation, cell migration, and vesicle and organelle transport. As such, they have been extensively studied at multiple levels of resolution (e.g., from structural biology to cell biology). Despite these efforts, there remain significant gaps in our knowledge concerning how microtubule-binding proteins bind to microtubules, how dynamics connect different conformational states, and how these interactions and dynamics affect cellular processes. Structures of microtubule-associated proteins assembled on polymeric microtubules are not known at atomic resolution. Here, we report a structure of the cytoskeleton-associated protein glycine-rich (CAP-Gly) domain of dynactin motor on polymeric microtubules, solved by magic angle spinning NMR spectroscopy. We present the intermolecular interface of CAP-Gly with microtubules, derived by recording direct dipolar contacts between CAP-Gly and tubulin using double rotational echo double resonance (dREDOR)-filtered experiments. Our results indicate that the structure adopted by CAP-Gly varies, particularly around its loop regions, permitting its interaction with multiple binding partners and with the microtubules. To our knowledge, this study reports the first atomic-resolution structure of a microtubule-associated protein on polymeric microtubules. Our approach lays the foundation for atomic-resolution structural analysis of other microtubule-associated motors.magic angle spinning NMR | microtubules | dynactin's CAP-Gly domain | structure determination | intermolecular interface determination

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Section: Resultssupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Yan

Guo

Hou

et al. 2015

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

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“…In the future, it will be interesting to study how p150 interacts with the microtubule in its GTP‐bound state as this is likely the relevant configuration for transport initiation. It will be interesting to understand how this interaction mode compares to the reported structure of the p150 CAP‐Gly domain and its neighbouring acidic region binding in a flexible manner to the C‐terminal tubulin tails of GDP/taxol microtubules (Wang et al , 2014). …”

Section: Discussionmentioning

confidence: 99%

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

et al. 2017

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Cytoplasmic dynein is involved in a multitude of essential cellular functions. Dynein's activity is controlled by the combinatorial action of several regulatory proteins. The molecular mechanism of this regulation is still poorly understood. Using purified proteins, we reconstitute the regulation of the human dynein complex by three prominent regulators on dynamic microtubules in the presence of end binding proteins (EBs). We find that dynein can be in biochemically and functionally distinct pools: either tracking dynamic microtubule plus‐ends in an EB‐dependent manner or moving processively towards minus ends in an adaptor protein‐dependent manner. Whereas both dynein pools share the dynactin complex, they have opposite preferences for binding other regulators, either the adaptor protein Bicaudal‐D2 (BicD2) or the multifunctional regulator Lissencephaly‐1 (Lis1). BicD2 and Lis1 together control the overall efficiency of motility initiation. Remarkably, dynactin can bias motility initiation locally from microtubule plus ends by autonomous plus‐end recognition. This bias is further enhanced by EBs and Lis1. Our study provides insight into the mechanism of dynein regulation by dissecting the distinct functional contributions of the individual members of a dynein regulatory network.

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“…Because the association of the CENP-E C-terminal kinetochore-binding domain with microtubules depends little (20% reduction) on tubulin C-terminal tails [370], these results suggest that microtubule detyrosination directly regulates recognition by the CENP-E motor domain. In contrast, the recognition of tyrosinated microtubules by Dynein has been shown to involve p150/Dynactin [358,360] and structural reconstructions have indicated that this interaction is mediated by the GKNDG motif on the CAP-Gly domain of p150/Dynactin [371,372]. …”

Section: Mechanisms Of Chromosome Congressionmentioning

confidence: 99%

Mechanisms of Chromosome Congression during Mitosis

Maiato

Gomes

Sousa

et al. 2017

Biology

170

176

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Chromosome congression during prometaphase culminates with the establishment of a metaphase plate, a hallmark of mitosis in metazoans. Classical views resulting from more than 100 years of research on this topic have attempted to explain chromosome congression based on the balance between opposing pulling and/or pushing forces that reach an equilibrium near the spindle equator. However, in mammalian cells, chromosome bi-orientation and force balance at kinetochores are not required for chromosome congression, whereas the mechanisms of chromosome congression are not necessarily involved in the maintenance of chromosome alignment after congression. Thus, chromosome congression and maintenance of alignment are determined by different principles. Moreover, it is now clear that not all chromosomes use the same mechanism for congressing to the spindle equator. Those chromosomes that are favorably positioned between both poles when the nuclear envelope breaks down use the so-called “direct congression” pathway in which chromosomes align after bi-orientation and the establishment of end-on kinetochore-microtubule attachments. This favors the balanced action of kinetochore pulling forces and polar ejection forces along chromosome arms that drive chromosome oscillatory movements during and after congression. The other pathway, which we call “peripheral congression”, is independent of end-on kinetochore microtubule-attachments and relies on the dominant and coordinated action of the kinetochore motors Dynein and Centromere Protein E (CENP-E) that mediate the lateral transport of peripheral chromosomes along microtubules, first towards the poles and subsequently towards the equator. How the opposite polarities of kinetochore motors are regulated in space and time to drive congression of peripheral chromosomes only now starts to be understood. This appears to be regulated by position-dependent phosphorylation of both Dynein and CENP-E and by spindle microtubule diversity by means of tubulin post-translational modifications. This so-called “tubulin code” might work as a navigation system that selectively guides kinetochore motors with opposite polarities along specific spindle microtubule populations, ultimately leading to the congression of peripheral chromosomes. We propose an integrated model of chromosome congression in mammalian cells that depends essentially on the following parameters: (1) chromosome position relative to the spindle poles after nuclear envelope breakdown; (2) establishment of stable end-on kinetochore-microtubule attachments and bi-orientation; (3) coordination between kinetochore- and arm-associated motors; and (4) spatial signatures associated with post-translational modifications of specific spindle microtubule populations. The physiological consequences of abnormal chromosome congression, as well as the therapeutic potential of inhibiting chromosome congression are also discussed.

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Structural basis for the extended CAP-Gly domains of p150 ^glued binding to microtubules and the implication for tubulin dynamics

Cited by 43 publications

References 34 publications

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

Mechanisms of Chromosome Congression during Mitosis

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Structural basis for the extended CAP-Gly domains of p150 glued binding to microtubules and the implication for tubulin dynamics

Cited by 43 publications

References 34 publications

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

Mechanisms of Chromosome Congression during Mitosis

Contact Info

Product

Resources

About

Structural basis for the extended CAP-Gly domains of p150 ^glued binding to microtubules and the implication for tubulin dynamics