title: Bacterial tubulin distinct loop sequences and primitive assembly properties support its origin from a eukaryotic tubulin ancestor.
abstract: The structure of the unique bacterial tubulin BtubA/B from Prosthecobacter is very similar to eukaryotic αβ-tubulin but, strikingly, BtubA/B fold without eukaryotic chaperones. Our sequence comparisons indicate that BtubA and BtubB do not really correspond to either α- or β-tubulin but have mosaic sequences with intertwining features from both. Their nucleotide-binding loops are more conserved, and their more divergent sequences correspond to discrete surface zones of tubulin involved in microtubule assembly and binding to eukaryotic cytosolic chaperonin, which is absent from the Prosthecobacter dejongeii draft genome. BtubA/B cooperatively assembles over a wider range of conditions than αβ-tubulin, forming pairs of protofilaments that coalesce into bundles instead of microtubules, and it lacks the ability to differentially interact with divalent cations and bind typical tubulin drugs. Assembled BtubA/B contain close to one bound GTP and GDP. Both BtubA and BtubB subunits hydrolyze GTP, leading to disassembly. The mutant BtubA/B-S144G in the tubulin signature motif GGG(T/S)G(S/T)G has strongly inhibited GTPase, but BtubA-T147G/B does not, suggesting that BtubB is a more active GTPase, like β-tubulin. BtubA/B chimera bearing the β-tubulin loops M, H1-S2, and S9-S10 in BtubB fold, assemble, and have reduced GTPase activity. However, introduction of the α-tubulin loop S9-S10 with its unique eight-residue insertion impaired folding. From the sequence analyses, its primitive assembly features, and the properties of the chimeras, we propose that BtubA/B were acquired shortly after duplication of a spontaneously folding α- and β-tubulin ancestor, possibly by horizontal gene transfer from a primitive eukaryotic cell, followed by divergent evolution.
authors.affiliation: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Martin-Galiano
authors.forename: María A
authors.initials: MA
authors.lastname: Oliva
authors.forename: Laura
authors.initials: L
authors.lastname: Sanz
authors.forename: Anamitra
authors.initials: A
authors.lastname: Bhattacharyya
authors.forename: Marina
authors.initials: M
authors.lastname: Serna
authors.forename: Hugo
authors.initials: H
authors.lastname: Yebenes
authors.forename: Jose M
authors.initials: JM
authors.lastname: Valpuesta
authors.forename: Jose M
authors.initials: JM
authors.lastname: Andreu
mesh_terms: D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D005057:Eukaryotic Cells / Q000502:physiology*; D019143:Evolution, Molecular; D022761:Gene Transfer, Horizontal / Q000502:physiology*; D006090:Gram-Negative Bacteria / Q000502:physiology*; D006153:Guanosine Diphosphate / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D017510:Protein Folding; D017433:Protein Structure, Secondary; D014404:Tubulin / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D014404:Tubulin; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate