Lattice defects induced by microtubule-stabilizing agents exert a long-range effect on microtubule growth by promoting catastrophes.
Lattice defects induced by microtubule-stabilizing agents exert a long-range effect on microtubule growth by promoting catastrophes.
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DOI：
10.1073/pnas.2112261118
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发表时间：
2021-12-21
影响因子：
11.1
通讯作者：
Akhmanova A
中科院分区：
文献类型：
--
作者：
Rai A;Liu T;Katrukha EA;Estévez-Gallego J;Manka SW;Paterson I;Díaz JF;Kapitein LC;Moores CA;Akhmanova A
微管是细胞分裂、生长和分化的主要细胞骨架丝。微管可以在生长和缩短阶段之间快速切换，这种动态行为对于微管阵列的形成至关重要。为了深入了解控制微管动力学的机制，在这里，我们使用了微管稳定剂，如紫杉醇及其荧光类似物来操纵微管原丝数量，并在微管晶格中产生稳定的缺陷，这些缺陷可以使用荧光显微镜观察到。我们发现，微管聚合速率增加与原丝数量和药物诱导的微管晶格的不连续性可以促进加端灾难在几微米的距离。我们的数据表明，在微管壁的结构缺陷可以有长距离的传播影响微管尖端动力学。微管是动态的细胞骨架聚合物，自发地在生长和收缩阶段之间切换。从生长到收缩的转变（称为灾难）的可能性随着微管年龄的增加而增加，但其潜在机制却知之甚少。在这里，我们着手测试聚合过程中形成的微管晶格缺陷是否会影响正端的生长。为了产生具有晶格缺陷的微管，我们使用微管稳定剂，其促进具有不同原丝数目的聚合物的形成。通过在稳定的微管种子的成核和随后的聚合阶段期间采用不同的试剂，我们可以重复地诱导原丝数目的开关并诱导稳定的晶格缺陷。这种药物诱导的缺陷导致了频繁的灾难，当微管在相同条件下生长但没有原丝数量错配时，没有观察到这种灾难。在缺陷部位切断微管足以抑制灾难。我们的结论是，微管晶格内的结构缺陷可以施加的影响，可以长距离传播，并影响微管端的动态状态。
Microtubules are major cytoskeletal filaments important for cell division, growth, and differentiation. Microtubules can rapidly switch between phases of growth and shortening, and this dynamic behavior is essential for shaping microtubule arrays. To obtain insights into mechanisms controlling microtubule dynamics, here we used microtubule-stabilizing agents such as Taxol and their fluorescent analogs to manipulate microtubule protofilament number and generate stable defects in microtubule lattices that can be visualized using fluorescence microscopy. We show that microtubule polymerization rate increases with protofilament number and that drug-induced microtubule lattice discontinuities can promote plus-end catastrophes at a distance of several micrometers. Our data indicate that structural defects in the microtubule wall can have long-range propagating effects on microtubule tip dynamics. Microtubules are dynamic cytoskeletal polymers that spontaneously switch between phases of growth and shrinkage. The probability of transitioning from growth to shrinkage, termed catastrophe, increases with microtubule age, but the underlying mechanisms are poorly understood. Here, we set out to test whether microtubule lattice defects formed during polymerization can affect growth at the plus end. To generate microtubules with lattice defects, we used microtubule-stabilizing agents that promote formation of polymers with different protofilament numbers. By employing different agents during nucleation of stable microtubule seeds and the subsequent polymerization phase, we could reproducibly induce switches in protofilament number and induce stable lattice defects. Such drug-induced defects led to frequent catastrophes, which were not observed when microtubules were grown in the same conditions but without a protofilament number mismatch. Microtubule severing at the site of the defect was sufficient to suppress catastrophes. We conclude that structural defects within the microtubule lattice can exert effects that can propagate over long distances and affect the dynamic state of the microtubule end.
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影响因子：
4
作者：
Bowne-Anderson, Hugo;Zanic, Marija;Kauer, Monika;Howard, Jonathon
通讯作者：
Howard, Jonathon
微流体辅助小管蛋白清洗探测的微管老化。
影响因子：
3.3
作者：
Duellberg C;Cade NI;Surrey T
通讯作者：
Surrey T
DOI：
10.1073/pnas.2001546117
发表时间：
2020-07-21
影响因子：
11.1
作者：
Debs, Garrett E.;Cha, Michael;Sindelar, Charles V.
通讯作者：
Sindelar, Charles V.
影响因子：
3.3
作者：
Chaaban S;Brouhard GJ
通讯作者：
Brouhard GJ
影响因子：
11.8
作者：
Aher A;Kok M;Sharma A;Rai A;Olieric N;Rodriguez-Garcia R;Katrukha EA;Weinert T;Olieric V;Kapitein LC;Steinmetz MO;Dogterom M;Akhmanova A
通讯作者：
Akhmanova A