Accessibility of the unstructured α-tubulin C-terminal tail is controlled by microtubule lattice conformation.

Abstract

Microtubules are cytoskeletal filaments that self-assemble from the protein tubulin, a heterodimer of α-tubulin and β-tubulin, and are important for cell mechanics, migration, and division. Much work has focused on how the nucleotide state of β-tubulin regulates the structure and dynamics of microtubules. In contrast, less is known about the structure and function of the C-terminal tails (CTTs) of α- and β-tubulin which are thought to freely protrude from the surface of the microtubule. To study the CTT of α-tubulin, we developed three different biosensors that bind the tyrosinated α-tubulin CTT (Y-αCTT). Surprisingly, live imaging of the probes indicates that the Y-αCTT is minimally accessible along the microtubule lattice under normal cellular conditions. Lattice binding of the Y-αCTT probes can be increased by three different ways of changing the tubulin conformational state: the drug Taxol, expression of microtubule-associated proteins (MAPs) that recognize or promote an expanded tubulin conformation, or expression of tubulin that cannot hydrolyze GTP. Molecular dynamics simulations indicate that the Y-αCTT undergoes numerous transient interactions with the bodies of α-tubulin and β-tubulin in the lattice, and that the frequency of these interactions is regulated by the tubulin nucleotide state. These findings suggest that accessibility of the Y-αCTT is locally governed by nucleotide- and MAP-dependent conformational changes to tubulin subunits within the microtubule lattice.