TitleGenetically encoded live-cell sensor for tyrosinated microtubules.
Publication TypeJournal Article
Year of Publication2020
AuthorsKesarwani S, Lama P, Chandra A, P Reddy P, Jijumon AS, Bodakuntla S, Rao BM, Janke C, Das R, Sirajuddin M
JournalJ Cell Biol
Volume219
Issue10
Date Published2020 Oct 05
ISSN1540-8140
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.

DOI10.1083/jcb.201912107
Alternate JournalJ Cell Biol
PubMed ID32886100
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