Genetically encoded live-cell sensor for tyrosinated microtubules.
Journal
The Journal of cell biology
ISSN: 1540-8140
Titre abrégé: J Cell Biol
Pays: United States
ID NLM: 0375356
Informations de publication
Date de publication:
05 10 2020
05 10 2020
Historique:
received:
19
12
2019
revised:
16
04
2020
accepted:
21
07
2020
entrez:
5
9
2020
pubmed:
5
9
2020
medline:
25
3
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32886100
pii: 152071
doi: 10.1083/jcb.201912107
pmc: PMC7659708
pii:
doi:
Substances chimiques
Microtubule-Associated Proteins
0
Tubulin
0
Tyrosine
42HK56048U
Vincristine
5J49Q6B70F
Nocodazole
SH1WY3R615
Colchicine
SML2Y3J35T
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : DBT-Wellcome Trust India Alliance
ID : IA/E/15/1/502339
Pays : India
Organisme : DBT-Wellcome Trust India Alliance
ID : IA/I/14/2/501533
Pays : India
Informations de copyright
© 2020 Kesarwani et al.
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