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Alemany, K.

Publications and source records attributed to Alemany, K..

2 recordsLinked to original sources

Role of α-tubulin helix 11' in heterodimer conformation and microtubule dynamics

Tubulin heterodimers transition from a curved conformation in solution to straight conformation when assembled into the microtubule lattice. Many proteins and small molecules alter microtubule dynamics by binding and stabilizing the curved or straight conformations; however, we have a poor understanding of the regions of the tubulin heterodimer that contribute to this transition and whether these conformations represent sources of phenotypic variation in human disease and across species. We previously identified a role for -tubulin helix 11 (H11) in the curved-to-straight transition. Using computational simulations and tubulin mutants in budding yeast, we show that H11 variants associated with human disease destabilize the curved conformation of the heterodimer and disrupt -tubulin function in cells. Comparing -tubulin sequences across eukaryotes demonstrates strong conservation of H11 with the exception of several -tubulin isotypes expressed during mitosis in the amoebae Naegleria fowleri and Naegleria gruberi. Introducing H11 sequence variants from Naegleria in budding yeast -tubulin increases heterodimer exchange at microtubule plus ends and destabilizes mitotic spindles. We provide evidence that interactions between -tubulin H11 and {beta}-tubulin H8 stabilize the curved conformation of tubulin heterodimers, and that the equilibrium between curved and straight conformations is an ancient feature of tubulin evolution.

cell biology↗

Kinesin-8/Kip3 requires β-tubulin tail for depolymerase activity

Carboxy-terminal tails (CTTs) of tubulin proteins are sites of regulating microtubule function. We previously conducted a genetic interaction screen and identified Kip3, a kinesin-8 motor, as potentially requiring the {beta}-tubulin CTT ({beta}-CTT) for function. Here we use budding yeast to define how {beta}-CTT promotes Kip3 function and the features of {beta}-CTT that are important for this mechanism. We find that {beta}-CTT is necessary for Kip3 depolymerase activity, but not for microtubule binding and motility. Mutant yeast cells lacking {beta}-CTT show increased accumulation of Kip3 at plus ends and along microtubules, but no increase in catastrophe when Kip3 is overexpressed. In vitro experiments show that the {beta}-CTT is necessary for Kip3 to form a tight complex with soluble tubulin but is unnecessary for Kip3 to bind tubulin in the microtubule lattice. These results suggest a model in which {beta}-CTT promotes Kip3 depolymerase activity by supporting a Kip3-tubulin binding state that is only accessible at the microtubule plus end or in solution.

cell biology↗