Human β-Tubulin Isotypes Can Regulate Microtubule Protofilament Number and Stability

Cell biological studies have shown that protofilament number, a fundamental feature of microtubules, can correlate with the expression of different tubulin isotypes. However, it is not known if tubulin isotypes directly control this basic microtubule property. Here, we report high-resolution cryo-EM reconstructions (3.5–3.65 Å) of purified human α1B/β3 and α1B/β2B microtubules and find that the β-tubulin isotype can determine protofilament number. Comparisons of atomic models of 13- and 14-protofilament microtubules reveal how tubulin subunit plasticity, manifested in “accordion-like” distributed structural changes, can accommodate distinct lattice organizations. Furthermore, compared to α1B/β3 microtubules, α1B/β2B filaments are more stable to passive disassembly and against depolymerization by MCAK or chTOG, microtubule-associated proteins with distinct mechanisms of action. Mixing tubulin isotypes in different proportions results in microtubules with protofilament numbers and stabilities intermediate to those of isotypically pure filaments. Together, our findings indicate that microtubule protofilament number and stability can be controlled through β-tubulin isotype composition. •Human β-tubulin isotypes can control microtubule protofilament number•3.5-Å cryo-EM structures of microtubules with human α1B/β2B- and α1B/β3-tubulin•α1B/β2B microtubules are intrinsically more stable than α1B/β3 filaments•β-tubulin isotypes can regulate MCAK- and chTOG-dependent depolymerization Microtubules are dynamic polymers of α/β-tubulin heterodimers that align head-to-tail to form protofilaments. Distinct α- and β-tubulin isotypes are expressed in different cell types to achieve diversity in microtubule organization and function. Ti et al. show that human β-tubulin isotypes can regulate basic microtubule properties such as protofilament number and stability.