Reviewer #1 (Public Review): Interface-acting nucleotide controls polymerization dynamics at microtubule plus- and minus-ends DOI Open Access
Lauren A McCormick, Joseph M. Cleary, William O. Hancock

и другие.

Опубликована: Окт. 30, 2023

GTP-tubulin is preferentially incorporated at growing microtubule ends, but the biochemical mechanism by which bound nucleotide regulates strength of tubulin:tubulin interactions debated. The 'self-acting' (cis) model posits that (GTP or GDP) to a particular tubulin dictates how strongly interacts, whereas 'interface-acting' (trans) interface two dimers determinant. We identified testable difference between these mechanisms using mixed simulations elongation: with self-acting nucleotide, plus- and minus-end growth rates decreased in same proportion amount GDP-tubulin, interface-acting plus-end disproportionately. then experimentally measured elongation nucleotides observed disproportionate effect GDP-tubulin on rates. Simulations were consistent binding 'poisoning' plus-ends not minus-ends. Quantitative agreement experiments required exchange terminal subunits mitigate poisoning there. Our results indicate interfacial determines interaction strength, thereby settling longstanding debate over state dynamics.

Язык: Английский

Structures, energetics, and dynamics of active tubulin self-organization DOI Creative Commons
Uri Raviv

Current Opinion in Solid State and Materials Science, Год журнала: 2025, Номер 36, С. 101219 - 101219

Опубликована: Март 16, 2025

Язык: Английский

Процитировано

1

Regulation of microtubule growth rates and their impact on chromosomal instability DOI
Lia Mara Gomes Paim, Susanne Bechstedt

Cell Cycle, Год журнала: 2025, Номер unknown, С. 1 - 20

Опубликована: Апрель 22, 2025

Microtubules are polymers of α/β tubulin dimers that build the mitotic spindle, which segregates duplicated chromosomes during cell division. Microtubule function is governed by dynamic instability, whereby cycles growth and shrinkage contribute to forces necessary for chromosome movement. Regulation microtubule velocity requires cycle-dependent changes in expression, localization activity microtubule-associated proteins (MAPs) as well post-translational modifications modulate dynamics. It has become clear optimal velocities required proper segregation ploidy maintenance. Suboptimal rates can result from altered MAPs could lead aneuploidy, possibly disrupting establishment bundles at kinetochores altering mechanical sister chromatid segregation. Future work using high-resolution, low-phototoxicity microscopy novel fluorescent markers will be invaluable obtaining deeper mechanistic insights into how processes

Язык: Английский

Процитировано

0

CLASPs stabilize the pre-catastrophe intermediate state between microtubule growth and shrinkage DOI Creative Commons
Elizabeth J. Lawrence, Saptarshi Chatterjee, Marija Žanić

и другие.

The Journal of Cell Biology, Год журнала: 2023, Номер 222(7)

Опубликована: Май 15, 2023

Cytoplasmic linker-associated proteins (CLASPs) regulate microtubules in fundamental cellular processes. CLASPs stabilize dynamic by suppressing microtubule catastrophe and promoting rescue, the switch-like transitions between growth shrinkage. How specifically modulate is not understood. Here, we investigate effects of on pre-catastrophe intermediate state dynamics, employing distinct substrates to mimic state. Surprisingly, find that CLASP1 promotes depolymerization stabilized presence GTP, but absence nucleotide. This activity also observed for CLASP2 family members a minimal TOG2-domain construct. Conversely, stabilizes unstable upon tubulin dilution GTP. Strikingly, our results reveal drives with vastly different inherent stabilities into same slowly depolymerizing nucleotide-dependent manner. We interpret this as Therefore, conclude suppress stabilizing

Язык: Английский

Процитировано

8

Heterogeneous local structures of the microtubule lattice revealed by cryo-ET and non-averaging analysis DOI Creative Commons
Hanjin Liu, Hiroshi Yamaguchi, Masahide Kikkawa

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Май 2, 2024

Summary Microtubule cytoskeletons play pivotal roles in various cellular processes, including cell division and locomotion, by dynamically changing their length distribution cells through tubulin polymerization/depolymerization. Recent structural studies have revealed the polymorphic lattice structure of microtubules closely correlate with microtubule dynamics, but were limited to averaged structures. To reveal transient localized structures, such as GTP-cap, we developed several non-averaging methods for cryogenic electron tomography precisely measure longitudinal spacing helical twisting individual lattices at subunit level. Our analysis that polymerizing depolymerizing ends share a similar regards spacing. The most distinctive property specific plus end was left-handed inter-dimer interface, suggesting might accelerate fast polymerization ends. uncovered heterogeneity native will be indispensable study dynamics under physiological contexts or during events.

Язык: Английский

Процитировано

2

Measuring and modeling forces generated by microtubules DOI
Nikita B. Gudimchuk, Veronika V. Alexandrova

Biophysical Reviews, Год журнала: 2023, Номер 15(5), С. 1095 - 1110

Опубликована: Окт. 1, 2023

Язык: Английский

Процитировано

5

CLASPs stabilize the intermediate state between microtubule growth and catastrophe DOI Creative Commons
Elizabeth J. Lawrence, Saptarshi Chatterjee, Marija Žanić

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2022, Номер unknown

Опубликована: Дек. 4, 2022

ABSTRACT CLASPs regulate microtubules in many fundamental cellular processes. stabilize dynamic by suppressing catastrophe and promoting rescue, the switch-like transitions between microtubule growth shrinkage. However, molecular mechanisms underlying CLASP’s activity are not understood. Here, we investigate effects of on distinct substrates absence tubulin to gain insight into how dynamics. Surprisingly, find that human CLASP1 depolymerizes stable presence GTP, but nucleotide. Conversely, stabilizes upon dilution GTP. Our results demonstrate drives with different inherent stabilities same slowly-depolymerizing state a nucleotide-dependent manner. We interpret this as pre-catastrophe intermediate Thus, conclude shrinkage suppress promote rescue.

Язык: Английский

Процитировано

8

Interface-acting nucleotide controls polymerization dynamics at microtubule plus- and minus-ends DOI Creative Commons
Lauren A McCormick, Joseph M. Cleary, William O. Hancock

и другие.

eLife, Год журнала: 2023, Номер 12

Опубликована: Июль 21, 2023

GTP-tubulin is preferentially incorporated at growing microtubule ends, but the biochemical mechanism by which bound nucleotide regulates strength of tubulin:tubulin interactions debated. The 'self-acting' (cis) model posits that (GTP or GDP) to a particular tubulin dictates how strongly interacts, whereas 'interface-acting' (trans) interface two dimers determinant. We identified testable difference between these mechanisms using mixed simulations elongation: with self-acting nucleotide, plus- and minus-end growth rates decreased in same proportion amount GDP-tubulin, interface-acting plus-end disproportionately. then experimentally measured elongation nucleotides observed disproportionate effect GDP-tubulin on rates. Simulations were consistent binding 'poisoning' plus-ends not minus-ends. Quantitative agreement experiments required exchange terminal subunits mitigate poisoning there. Our results indicate interfacial determines interaction strength, thereby settling longstanding debate over state dynamics.

Язык: Английский

Процитировано

4

Interface-acting nucleotide controls polymerization dynamics at microtubule plus- and minus-ends DOI Creative Commons
Lauren A McCormick, Joseph M. Cleary, William O. Hancock

и другие.

eLife, Год журнала: 2024, Номер 12

Опубликована: Янв. 5, 2024

GTP-tubulin is preferentially incorporated at growing microtubule ends, but the biochemical mechanism by which bound nucleotide regulates strength of tubulin:tubulin interactions debated. The ‘self-acting’ (cis) model posits that (GTP or GDP) to a particular tubulin dictates how strongly interacts, whereas ‘interface-acting’ (trans) interface two dimers determinant. We identified testable difference between these mechanisms using mixed simulations elongation: with self-acting nucleotide, plus- and minus-end growth rates decreased in same proportion amount GDP-tubulin, interface-acting plus-end disproportionately. then experimentally measured elongation nucleotides observed disproportionate effect GDP-tubulin on rates. Simulations were consistent binding ‘poisoning’ plus-ends not minus-ends. Quantitative agreement experiments required exchange terminal subunits mitigate poisoning there. Our results indicate interfacial determines interaction strength, thereby settling longstanding debate over state dynamics.

Язык: Английский

Процитировано

1

Modulation of Microtubule Dynamics by Monovalent Ions DOI Creative Commons
Simon Fernandes, Charlotte Aumeier

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 26, 2024

ABSTRACT The microtubule cytoskeleton is a dynamic network essential for many cellular processes, influenced by physicochemical factor such as temperature, pH, dimer concentration and ionic environment. In this study, we used in vitro reconstitution assays to examine the effects of four monovalent ions (Na + , K Cl - Ac ) on dynamics, uncovering distinct each ion. Na was found increase dynamicity raising catastrophe frequency, polymerization depolymerization speeds, ultimately reducing lifetime 80 %. Conversely, boosts nucleation stabilizes microtubules increasing rescue frequency preventing breakages, resulting longer with extended lifetimes. appeared potentiate while had minimal impact parameters. These findings demonstrate that have opposing destabilizing stabilizing structure. This mainly through modulation tubulin-tubulin interactions rather than affecting hydrolysis rate. conclusion, ion identity plays crucial role modulating dynamics. Understanding environment microtubule-related research, it significantly influences behavior, stability, other proteins. SIGNIFICANCE STATEMENT vital processes concentration, how these factors regulate elucidating dynamics stability. Our reveal increases %, enhances microtubules. potentiates has impact. highlight crucially modulates influencing stability interactions.

Язык: Английский

Процитировано

0

EB3-informed dynamics of the microtubule stabilizing cap during stalled growth DOI Creative Commons
Maurits Kok, Florian Huber,

Svenja-Marei Kalisch

и другие.

Biophysical Journal, Год журнала: 2024, Номер 124(2), С. 227 - 244

Опубликована: Ноя. 27, 2024

Язык: Английский

Процитировано

0