Structural insight into the stabilization of microtubules by taxanes

Prota, A.E.; Lucena‐Agell, Daniel; Ma, Yuntao; Estévez-Gallego, Juan; Roca, Carlos P.; Josa-Prado, Fernando; Goossens, Kenneth; Giménez-Abián, Juan F.; Li, Shuo; Canales, Ángeles; Bargsten, Katja; Andreu, José Manuel; Altmann, Karl‐Heinz; Olieric, Natacha; Kamimura, Shinji; Mühlethaler, Tobias; Oliva, M.A.; Steinmetz, Michel O.; Fang, Wei‐Shuo; Díaz, J. Fernando

doi:10.1101/2021.07.20.453061

Cited by 3 publications

(5 citation statements)

References 86 publications

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“…It is already known that effector-dependent stabilisation and effector-dependent expansion of the microtubule lattice are not necessarily coupled. Prota et al 35 recently showed that baccatin III, a taxol precursor, expands but only marginally stabilises brain GDP-microtubules.…”

Section: Discussionmentioning

confidence: 99%

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Taxol acts differently on different tubulin isotypes

Chew

Cross

2023

Preprint

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Taxol is a critically important cancer drug that stabilises microtubules. We report that taxol acts differently on different metazoan tubulin isotypes. 50 nM taxol blocks catastrophe of human or zebrafish α1β4 but has no effect on human α1β3 microtubules. 500 nM taxol blocks catastrophe in both α1β3 and α1β4 microtubules but introduces kinks only into α1β4 microtubules. Taxol washout relaxes the kinks, suggesting taxol expands α1β4 but not α1β3 lattices. Kinesin-driven microtubule gliding detects this conformational shift - α1β4 microtubules glide at ~450 nm/sec in 400 nM taxol, but at ~750 nm/sec in 10 μM taxol, whereas α1β3 microtubules glide at ~450 nm/sec, even in 10 μM taxol. Thus, taxol readily stabilises α1β4 GDP-tubulin lattices and shifts them to a fast-gliding conformation, but stabilises α1β3 lattices much less readily and without shifting their conformation. These isotype-specific actions of taxol may drive the switch to β3 tubulin commonly seen in taxol-resistant tumours.

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Section: Discussionmentioning

confidence: 99%

“…Recent work indicates that taxol binds the GDP-tubulin lattice much more tightly than it binds to free tubulin because the polymerisation switch 38 reconfigures the M-loop, which otherwise occludes the taxol site 35 . The lateral contacts made by the M-loops may be the dominant determinant of lattice stability 39 .…”

Section: Discussionmentioning

confidence: 99%

Taxol acts differently on different tubulin isotypes

Chew

Cross

2023

Preprint

View full text Add to dashboard Cite

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“…Recent work indicates that taxol binds the GDP-tubulin lattice much more tightly than it binds to free tubulin because the polymerisation switch 38 recon gures the M-loop, which otherwise occludes the taxol site 35 . The lateral contacts made by the M-loops may be the dominant determinant of lattice stability 39 .…”

Section: Discussionmentioning

confidence: 99%

“…It is already known that effector-dependent stabilisation and effector-driven expansion of the microtubule lattice are not necessarily coupled. Prota et al 35 recently showed that baccatin III, a taxol precursor, expands but only very marginally stabilises brain GDP-microtubules.…”

Section: Discussionmentioning

confidence: 99%

Taxol acts differently on different tubulin isotypes

Chew

Cross

2023

Preprint

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Taxol is a small molecule effector that allosterically locks tubulin into the microtubule lattice. We report that taxol has different effects on different single isotype microtubule lattices. Using in vitro reconstitution, we show that α1β4 human and zebrafish GDP-tubulin lattices are stabilised and expanded by taxol, whereas α1β3 human GDP-tubulin lattices require tenfold more taxol for stability and are not expanded. In kinesin motility assays, this isotype-specific mechanical action of taxol causes segmented isotype microtubules to deviate into loops, because the expanded β4 and compacted β3 segments try to glide at different rates. To explain, we propose that taxol switches GDP-microtubules into one of two different lattice-mechanical states, a compacted/slow-gliding state or an expanded/fast-gliding state, with switching to the expanded/fast-gliding state dependant on taxol occupancy and available to only some tubulin isotypes. In mixed isotype lattices, we find evidence that this lattice-mechanical switching occurs cooperatively.

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“…Microtubules are hollow polymers made of tubulin heterodimers, organised in a 'head to tail' fashion in the microtubule wall 98 . Taxol's core structure, baccatin III, binds to these curved tubulin molecules to elicit its effect 99 . The microtubule cytoskeleton becomes reorganised, forming large bundles.…”

Section: Paclitaxel (Taxol)mentioning

confidence: 99%

'I get by with a little help from my friends': The role of stromal cells in the breast cancer chemoresistance mechanism

Thompson¹

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Breast cancer is the most common cancer that affects women worldwide, and in New Zealand accounts for 1/3 of new cancer cases. The tumour microenvironment (TME) has an important role in how breast cancer cells respond to chemotherapeutic stress. Mesenchymal stromal cells (MStCs), a component of the TME, have been shown to function in the development of breast cancer cell chemoresistance, promoting survival and more aggressive cancer cell phenotypes. The effect of co-culture on the 4T1 murine breast cancer cell response to chemotherapy has been explored with DNA-damaging therapies but was yet to be explored against other chemotherapeutic mechanisms. Fluorescent-4T1 cells were co-cultured with compact bone-derived MStCs and treated with chemotherapies of differing modalities. Flow cytometry, microscopy and gene expression assays were used to assess the effect of co-culture on the 4T1 response to treatment with clinically relevant chemotherapy drugs. Fluorescent-4T1s had altered morphology, disrupted cell cycles, and decreased viability in response to drug treatment. MStCs had low positive expression of classical stromal markers and had reduced effects of chemotherapy compared to 4T1s. Co-culture with MStCs decreased the effect of drug on 4T1 viability, morphology and cell cycle progression. Increased contact between 4T1s and MStCs resulted in enhanced survival of 4T1 cells under chemotherapeutic stress. These results indicated that drug efficacy is decreased when cancer cells are in co-culture with supporting cells of the TME, independent of the modality of chemotherapeutic stress. This emphasises the importance of the TME to the cancer cell stress response and begins to delve into potential mechanisms for this enhanced interaction. Future research should investigate candidates for inhibiting this interaction between stromal and cancer cells.

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Structural insight into the stabilization of microtubules by taxanes

Cited by 3 publications

References 86 publications

Taxol acts differently on different tubulin isotypes

Taxol acts differently on different tubulin isotypes

Taxol acts differently on different tubulin isotypes

'I get by with a little help from my friends': The role of stromal cells in the breast cancer chemoresistance mechanism

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