Effects of osmotic force and torque on microtubule bundling and pattern formation.
ABSTRACT We report effects of polyethylene glycol (PEG, molecular weight of 35 kDa ) on microtubule (MT) bundling and pattern formation. Without PEG, polymerizing tubulin solutions of a few mg/ml that are initially subjected to a field that aligns MTs can spontaneously form striated birefringence patterns. These patterns form through MT alignment, bundling, and coordinated bundle buckling. With increasing PEG concentrations, solutions form progressively weaker patterns. At a sufficiently high PEG concentration ( approximately 0.5% by weight), the samples maintain a nearly uniform birefringence (i.e., no pattern) and laterally contract at a later stage. Concomitantly, on a microscopic level, the network of dispersed MTs that accompany the bundles in pure solutions disappear and the bundles become more distinct. We attribute the weakening of the pattern to the loss of the dispersed MT network, which is required to mediate the coordination of bundle buckling. We propose that the loss of the dispersed network and the enhanced bundling result from PEG associated osmotic forces that drive MTs together and osmotic torques that facilitate their bundling. Similarly, we attribute the lateral contraction of the samples to osmotic torques that tend to align crossing bundles in the network.
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ABSTRACT: Multivalent counterions can induce an effective attraction between like-charged rodlike polyelectrolytes, leading to the formation of polyelectrolyte bundles. In this paper, we calculate the equilibrium bundle size using a simple model in which the attraction between polyelectrolytes (assumed to be pairwise additive) is treated phenomenologically. If the counterions are pointlike, they almost completely neutralize the charge of the bundle, and the equilibrium bundle size diverges. When the counterions are large, however, steric and short-range electrostatic interactions prevent charge neutralization of the bundle, thus forcing the equilibrium bundle size to be finite. We also show that if the attractive interactions between the rods become frustrated as the bundle grows, finite-size bundles can be obtained with pointlike counterions.Physical Review E 07/2005; 71(6 Pt 1):060801. · 2.31 Impact Factor
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ABSTRACT: Microtubules (MTs) are hollow cylindrical polymers composed of alphabeta-tubulin heterodimers that align head-to-tail in the MT wall, forming linear protofilaments that interact laterally. We introduce a probe of the interprotofilament interactions within MTs and show that this technique gives insight into the mechanisms by which MT-associated proteins (MAPs) and taxol stabilize MTs. In addition, we present further measurements of the mechanical properties of MT walls, MT-MT interactions, and the entry of polymers into the MT lumen. These results are obtained from a synchrotron small angle x-ray diffraction (SAXRD) study of MTs under osmotic stress. Above a critical osmotic pressure, P(cr), we observe rectangular bundles of MTs whose cross sections have buckled to a noncircular shape; further increases in pressure continue to distort MTs elastically. The P(cr) of approximately 600 Pa provides, for the first time, a measure of the bending modulus of the interprotofilament bond within an MT. The presence of neuronal MAPs greatly increases P(cr), whereas surprisingly, the cancer chemotherapeutic drug taxol, which suppresses MT dynamics and inhibits MT depolymerization, does not affect the interprotofilament interactions. This SAXRD-osmotic stress technique, which has enabled measurements of the mechanical properties of MTs, should find broad application for studying interactions between MTs and of MTs with MAPs and MT-associated drugs.Biophysical Journal 12/2005; 89(5):3410-23. · 3.67 Impact Factor
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ABSTRACT: We present a model for the spontaneous formation of a striated pattern in polymerizing microtubule solutions. It describes the buckling of a single microtubule (MT) bundle within an elastic network formed by other similarly aligned and buckling bundles and unaligned MTs. Phase contrast and polarization microscopy studies of the temporal evolution of the pattern imply that the polymerization of MTs within the bundles creates the driving compressional force. Using the measured rate of buckling, the established MT force-velocity curve and the pattern wavelength, we obtain reasonable estimates for the MT bundle bending rigidity and the elastic constant of the network. The analysis implies that the bundles buckle as solid rods.Physical Review Letters 06/2007; 98(19):198103. · 7.94 Impact Factor