Year: 2020
Type: article
Abstract: Microtubule cytoskeleton exists in various biochemical forms in different cells due to tubulin posttranslational modifications (PTMs). Tubulin PTMs are known to affect microtubule stability, dynamics, and interaction with MAPs and motors in a specific manner, widely known as tubulin code hypothesis. At present, there exists no tool that can specifically mark tubulin PTMs in living cells, thus severely limiting our understanding of their dynamics and cellular functions. Using a yeast display library, we identified a binder against terminal tyrosine of α-tubulin, a unique PTM site. Extensive characterization validates the robustness and nonperturbing nature of our binder as tyrosination sensor, a live-cell tubulin nanobody specific towards tyrosinated microtubules. Using this sensor, we followed nocodazole-, colchicine-, and vincristine-induced depolymerization events of tyrosinated microtubules in real time and found each distinctly perturbs the microtubule polymer. Together, our work describes a novel tyrosination sensor and its potential applications to study the dynamics of microtubule and their PTM processes in living cells.
Source: The Journal of Cell Biology
Institutions: Manipal Academy of Higher Education, Institute for Stem Cell Biology and Regenerative Medicine, Tata Institute of Fundamental Research, National Centre for Biological Sciences, Université Paris-Sud, Centre National de la Recherche Scientifique, Université Paris Sciences et Lettres, Université Paris-Saclay, Intégrité du génome, ARN et cancer, Institut Curie, North Carolina State University
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: hybrid
Funders: European Molecular Biology Organization, Wellcome Trust, Children's Investment Fund Foundation, European Commission, Tata Institute of Fundamental Research, Department of Biotechnology, Ministry of Science and Technology, India, Agence Nationale de la Recherche, Indo-French Centre for the Promotion of Advanced Research, Fondation pour la Recherche Médicale, Institut National Du Cancer, The Wellcome Trust DBT India Alliance, Institut Curie, National Centre for Biological Sciences, Horizon 2020
Awards: A multidisciplinary approach to cell division: From human oocyte to synthetic biology, G1937183540, The tubulin code as mechanism for context-dependent fine-tuning of microtubule functions, G2180428777, G2993143320, G3058230636, Structure and function studies of sarcomere proteins implicated in cardiomyopathies, G346652276, G434337789, Modulation of physiological and oncogenic Ras protein signaling via plasma membrane clustering, G6539978045, G6803667113, G7427359084, G8066969849
OpenAlex ID: W3083369693