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Regulation of microtubule assembly in cultured fibroblasts.

ABSTRACT: Microtubule assembly in diploid human skin fibroblasts was studied by [3H]colchicine binding to disaggregated microtubule subunits and to total cell tubulin. Microtubule content per milligram of cell protein was critically dependent upon cell density. As cultures neared confluence, microtubules increased and total cell tubulin decreased; the percent of tubulin assembled into microtubules increased from 5.3 in spare cultures to 58.3 in confluent cultures. Microtubules disappeared with a half-time of 2 min in response to 0 degree C incubation and reformed upon rewarming. Brief treatment of intact cells with concanavalin A or cytochalasin A depolymerized microtubules to 55 or 56% of control levels. The effect of concanavalin A was prevented by alpha-methylmannoside. Fibroblast microtubule assembly was not significantly altered by cyclic nucleotides, ascorbate, glucose, insulin, medium calcium concentration, or calcium ionophore A23187.

SUBMITTER: Ostlund RE

PROVIDER: S-EPMC2110629 | biostudies-literature | 1980 May

REPOSITORIES: biostudies-literature

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Regulation of microtubule assembly in cultured fibroblasts.

Ostlund R E RE Leung J T JT Hajek S V SV

The Journal of cell biology 19800501 2

Microtubule assembly in diploid human skin fibroblasts was studied by [3H]colchicine binding to disaggregated microtubule subunits and to total cell tubulin. Microtubule content per milligram of cell protein was critically dependent upon cell density. As cultures neared confluence, microtubules increased and total cell tubulin decreased; the percent of tubulin assembled into microtubules increased from 5.3 in spare cultures to 58.3 in confluent cultures. Microtubules disappeared with a half-time ...[more]

PMID: 7372712

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Project description:Living systems utilize sophisticated biochemical regulators and various signal transduction mechanisms to program bio-molecular assemblies and their associated functions. Creating synthetic assemblies that can replicate the functional and signal-responsive properties of these regulators, while also interfacing with biomolecules, holds significant interest within the realms of supramolecular chemistry and chemical biology. This pursuit not only aids in understanding the fundamental design principles of life but also introduces novel capabilities that contribute to the advancements in medical and therapeutic research. In this study, we present a cucurbit[7]uril (CB[7]) host-guest system designed to regulate the dynamics and functions of microtubules (MTs) in living cells. To establish communication between MTs and CB[7] and to reversibly control MT function through host-guest recognition, we synthesized a two-faced docetaxel-p-xylenediamine (Xyl-DTX) derivative. While Xyl-DTX effectively stabilized polymerized MTs, inducing MT bundling and reducing dynamics in GFP-α-tubulin expressing cells, we observed a significant reduction in its MT-targeted activity upon threading with CB[7]. Leveraging the reversible nature of the host-guest complexation, we strategically reactivated the MT stabilizing effect by programming the guest displacement reaction from the CB[7]·Xyl-DTX complex using a suitable chemical signal, namely a high-affinity guest. This host-guest switch was further integrated into various guest activation networks, enabling 'user-defined' regulatory control over MT function. For instance, we demonstrated programmable control over MT function through an optical signal by interfacing it with a photochemical guest activation network. Finally, we showcased the versatility of this supramolecular system in nanotechnology-based therapeutic approaches, where a self-assembled nanoparticle system was employed to trigger the MT-targeted therapeutic effect from the CB[7]·Xyl-DTX complex.

Dataset Information

Regulation of microtubule assembly in cultured fibroblasts.

Publications

Regulation of microtubule assembly in cultured fibroblasts.

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