Microtubule dynamic instability: the role of cracks between protofilaments

Microtubules (MTs) are cytoplasmic protein polymers that are essential for fundamental cellular processes including the maintenance of cell shape, organelle transport and formation of the mitotic spindle. Microtubule dynamic instability is critical for these processes, but it remains poorly understood, in part because the relationship between the structure of the MT tip and the growth/depolymerization transitions is enigmatic. In previous work, we used computational models of dynamic instability to provide evidence that cracks (laterally unbonded regions) between protofilaments play a key role in the regulation of dynamic instability. Here we use computational models to investigate the connection between cracks and dynamic instability in more detail. Our work indicates that while cracks contribute to dynamic instability in a fundamental way, it is not the depth of the cracks per se that governs MT dynamic instability. Instead, what matters more is whether the cracks terminate in GTP-rich or GDP-rich regions of the MT. Based on these observations, we suggest that a functional "GTP cap" (i.e., one capable of promoting MT growth) is one where the cracks terminate in pairs of GTP-bound subunits, and that the likelihood of catastrophe rises significantly with the fraction of crack-terminating subunits that contain GDP. In addition to helping clarify the mechanism of dynamic instability, this idea could also explain how MT stabilizers work: proteins that introduce lateral cross-links between protofilaments would produce islands of GDP-bound tubulin that mimic GTP-rich regions in having strong lateral bonds, thus reducing crack propagation, suppressing catastrophe and promoting rescue.