Zehr, E. A. | bioRxiv Science

bioRxiv2023

Cryo-EM structures of human α1B/βI+βIVb microtubules shed light on isoform specific assembly

Microtubules are dynamic polymers assembled from {beta}-tubulin dimers. Mammals have multiple and {beta}-tubulin isoforms. Despite a high degree of conservation, microtubules assembled from different tubulin isoforms have unique dynamic properties. How isoform sequence variation affects polymerization interfaces and nucleotide dependent conformational changes in microtubules is still not well understood. Here we report 2.9[A] resolution cryo-EM structures of human 1B/{beta}I+{beta}IVb microtubules in the GDP state, and a GTP-like state, bound to GMPCPP. Our structures show that, similar to microtubules assembled from other mammalian isoforms, transition from the GTP to the GDP states in 1B/{beta}I+{beta}IVb microtubules results in strengthening of the longitudinal interface and an overall compaction of the axial dimer repeat distance in the lattice. Interestingly, we find that -tail residues link longitudinally adjacent tubulin dimers through interactions with two conserved arginine residues in {beta}-tubulin that are mutated in human disease. Comparative analysis of tubulin isoforms shows minimal isoform-specific effects at the longitudinal interface or the -tubulin lateral interface, but a high concentration of sequence variability in the second shell of residues away from the {beta}-tubulin lateral interface which can modulate polymerization interfaces and thus impact microtubule dynamics.

biophysics↗

bioRxiv2018

Mechanistic Origins of Dynamic Instability in Filaments from the Phage Tubulin, PhuZ

A bacteriophage-encoded tubulin homologue, PhuZ, harnesses dynamic instability to position genomes of KZ-like bacteriophage at the midline of their Pseudomonas hosts, facilitating phage infectivity. While much has been learned about molecular origins of microtubule dynamics, how GTP binding and hydrolysis control dynamics in the divergent 3-stranded PhuZ filaments is not understood. Here we present cryo-EM reconstructions of the PhuZ filamentin a pre-hydrolysis (3.5[A]) and three post-hydrolysis states (4.2 [A], 7.3 [A] and 8.1 [A] resolutions), likely representing distinct depolymerization stages. Core polymerization-induced structural changes reveal similarities to {beta}-tubulin, suggesting broad conservation within the tubulin family. By contrast, GTP hydrolysis is sensed quite differently and is communicated by the divergent PhuZ C-terminus to the lateral interface, leading to PhuZ polymer destabilization. This provides a contrasting molecular description of how nucleotide state can be harnessed by the tubulin fold to regulate filament assembly, metastability and disassembly.

cell biology↗