High-Resolution Model of the Microtubule

Nogales, Eva; Whittaker, Michael; Milligan, Ronald A.; Downing, Kenneth H.

doi:10.1016/s0092-8674(00)80961-7

Cited by 1,082 publications

(1,136 citation statements)

References 38 publications

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“…This particular finding is discussed in more detail below. This is unexpected since on cryo-images made in the absence of decoration the two tubulin subunit types are almost indistinguishable at this resolution (Nogales et al, 1999).…”

Section: Bereitgestellt Von | Universitaetsbibliothek Der Lmu Muenchenmentioning

confidence: 88%

“…The availability of atomic resolution X-ray structures of monomeric and dimeric kinesin Sack et al, 1997;Kozielski et al, 1997) and ncd (Sablin et al, , 1998 in combination with the near-atomic resolution EM-structure of zinc-induced tubulin sheets (Nogales et al, 1998) now provide detailed pictures of the microtubule structure (Nogales et al, 1999) and of the kinesin-tubulin complex (Hoenger et al, 2000;Kikkawa et al, 2000). There is now a general consensus on the tubulin interaction and binding geometry of monomeric kinesin and ncd (Hirose et al, 1995;Hoenger et al, 1995;Kikkawa et al, 1995;Arnal et al 1996;Hoenger and Milligan, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Hoenger

Doerhoefer²,

Woehlke³

et al. 2000

Biological Chemistry

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The surface topography of opened-up microtubule walls (sheets) decorated with monomeric and dimeric kinesin motor domains was investigated by freezedrying and unidirectional metal shadowing. Electron microscopy of surface-shadowed specimens produces images with a high signal/noise ratio, which enable a direct observation of surface features below 2 nm detail. Here we investigate the inner and outer surface of microtubules and tubulin sheets with and without decoration by kinesin motor domains. Tubulin sheets are flattened walls of microtubules, keeping lateral protofilament contacts intact. Surface shadowing reveals the following features: (i) when the microtubule outside is exposed the surface relief is dominated by the bound motor domains. Monomeric motor constructs generate a strong 8 nm periodicity, corresponding to the binding of one motor domain per ␣-␤-tubulin heterodimer. This surface periodicity largely disappears when dimeric kinesin motor domains are used for decoration, even though it is still visible in negatively stained or frozen hydrated specimens. This could be explained by disorder in the binding of the second (loosely tethered) kinesin head, and/or disorder in the coiled-coil tail. (ii) Both surfaces of undecorated sheets or microtubules, as well as the inner surface of decorated sheets, reveal a strong 4 nm repeat (due to the periodicity of tubulin monomers) and a weak 8 nm repeat (due to slight differences between ␣-and ␤-tubulin). The differences between ␣-and ␤-tubulin on the inner surface are stronger than expected from cryo-electron microscopy of unstained microtubules, indicating the existence of tubulin subdomain-specific surface properties that reflect the surface corrugation and hence metal deposition during evaporation. The 16 nm periodicity visible in some negatively stained specimens (caused by the pairing of cooperatively bound kinesin dimers) is not detected by surface shadowing.

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Section: Bereitgestellt Von | Universitaetsbibliothek Der Lmu Muenchenmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Hoenger

Doerhoefer²,

Woehlke³

et al. 2000

Biological Chemistry

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“…Flexibility of microtubules connected with the fact that microtubule subunits are bound more strongly inter-protofilaments than within-protofilaments in vitro was shown by many authors (Chrétien et al 1998;Nogales 1999;Nogales et al 1999;Löwe et al 2001;Meurer-Grob et al 2001;Kis et al 2002;Sept et al 2003;Krebs et al 2005;Tuszyński et al 2005;Wang and Nogales 2005;Nogales and Wang 2006;Drabik et al 2007;Hunyadi and Jánosi 2007).…”

Section: Discussionmentioning

confidence: 91%

Immunogold method evidences that kinesin and myosin bind to and couple microtubules and actin filaments in lipotubuloids of Ornithogalum umbellatum ovary epidermis

et al. 2013

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Lipotubuloids in ovary epidermis of Ornithogalum umbellatum which are a domain of cytoplasm containing a lot of lipid bodies, microtubules and actin filaments, ribosomes, endoplasmic reticulum as well as scarce mitochondria, microbodies, dictyosomes, autolytic vacuoles, exhibit progressive-rotary motion. The immunogold method demonstrated that microtubules and actin filaments of lipotubuloids might be connected with one another by myosin and kinesin. It was supposed that collaboration of motor proteins with actin filaments and microtubules makes autonomic high peripheral speed rotary motion of lipotubuloids in epidermis cells possible. Moreover, myosin was also detected in Golgi bodies in lipotubuloid. In lipotubuloids, the immunogold method demonstrated immunosignals after the use of an antibody to dynein light chains but spectroscopy mass analysis showed that in O. umbellatum epidermis lacked dynein heavy chains.

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“…1A). High resolution modeling of the tubulin oligomer within the low resolution ab initio shape resulted in a linear combination of three ␣␤-dimers similar to that seen in the tubulin protofilament (PDBcode 1TUB) (Nogales et al, 1998(Nogales et al, , 1999. The oligomer is not fully straight, with the terminal dimer slightly bent relative to the axis of the preceding linear tetramer (Fig.…”

Section: Saxs Analysis Of the Tubulin-ataxin-3 Complexmentioning

confidence: 86%

Interactions of ataxin-3 with its molecular partners in the protein machinery that sorts protein aggregates to the aggresome

Bonanomi

Mazzucchelli

D’Urzo

et al. 2014

The International Journal of Biochemistry & Cell Biology

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a b s t r a c tAtaxin-3 (AT3) is the protein that triggers the inherited neurodegenerative disorder spinocerebellar ataxia type 3 when its polyglutamine (polyQ) stretch close to the C-terminus exceeds a critical length. AT3 consists of the N-terminal globular Josephin domain (JD) and the C-terminal disordered one. It cleaves isopeptide bonds between ubiquitin monomers, an event involved in protein quality control mechanisms. AT3 has been implicated in the pathway that sorts aggregated protein to aggresomes via microtubules, in which dynein and histone deacetylase 6 (HDAC6) also seem to be involved. By taking advantage of small angle X-ray scattering (SAXS) and surface plasmon resonance (SPR), we have investigated the interaction of AT3 with tubulin and HDAC6. Based on SAXS results, the AT3 oligomer, consisting of 6-7 subunits, tightly binds to the tubulin hexameric oligomer in a "parallel" fashion. By SPR analysis we have demonstrated that AT3 binds to tubulin dimer with a 50 nM affinity. Binding fits with a Langmuir 1:1 model and involves a single binding interface. Nevertheless, the interaction surface consists of three distinct, discontinuous tubulin-binding regions (TBR), one located in the JD, and the two others in the disordered domain, upstream and downstream of the polyQ stretch. In the absence of any of the three TBRs, the affinity is drastically reduced. By SPR we have also provided the first evidence of direct binding of AT3 to HDAC6, with affinity in the range 0.1-1 M. These results shed light on the interactions among the components of the transport machinery that sorts aggregate protein to the aggresome, and pave the way to in vivo studies aimed at further clarifying their roles.

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High-Resolution Model of the Microtubule

Cited by 1,082 publications

References 38 publications

Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Immunogold method evidences that kinesin and myosin bind to and couple microtubules and actin filaments in lipotubuloids of Ornithogalum umbellatum ovary epidermis

Interactions of ataxin-3 with its molecular partners in the protein machinery that sorts protein aggregates to the aggresome

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