Fine structure and dynamics of EB3 binding zones on microtubules in fibroblast cells

V. V. Mustyatsa, A. V. Kostarev, A. V. Tvorogova, F. I. Ataullakhanov, N. B. Gudimchuk, Ivan Vorobyev

Research output: Contribution to journalArticle

Abstract

End-binding (EB) proteins associate with the growing tips of microtubules (MTs)and modulate their dynamics directly and indirectly, by recruiting essential factors to fine-tune MTs for their many essential roles in cells. Previously EB proteins have been shown to recognize a stabilizing GTP/GDP-Pi cap at the tip of growing MTs, but information about additional EB-binding zones on MTs has been limited. In this work, we studied fluorescence intensity profiles of one of the three mammalian EB-proteins, EB3, fused with red fluorescent protein (RFP). The distribution of EB3 on MTs in mouse fibroblasts frequently deviated from single exponential decay and exhibited secondary peaks. Those secondary peaks, which we refer to as EB3-islands, were detected on 56% comets of growing MTs and were encountered once per 44 s of EB3-RFP comet growth time with about 5 s half-lifetime. The majority of EB3-islands in the vicinity of MT tips was stationary and originated from EB3 comets moving with the growing MT tips. Computational modeling of the decoration of dynamic MT tips by EB3 suggested that the EB3-islands could not be explained simply by a stochastic first-order GTP hydrolysis/phosphate release. We speculate that additional protein factors contribute to EB3 residence time on MTs in cells, likely affecting MT dynamics.

Original languageEnglish
Pages (from-to)2105-2114
Number of pages10
JournalMolecular biology of the cell
Volume30
Issue number17
DOIs
Publication statusPublished - Aug 1 2019

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Microtubules
Fibroblasts
Islands
Carrier Proteins
Guanosine Triphosphate
Hydrolysis
Fluorescence
Phosphates

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Mustyatsa, V. V., Kostarev, A. V., Tvorogova, A. V., Ataullakhanov, F. I., Gudimchuk, N. B., & Vorobyev, I. (2019). Fine structure and dynamics of EB3 binding zones on microtubules in fibroblast cells. Molecular biology of the cell, 30(17), 2105-2114. https://doi.org/10.1091/mbc.E18-11-0723

Fine structure and dynamics of EB3 binding zones on microtubules in fibroblast cells. / Mustyatsa, V. V.; Kostarev, A. V.; Tvorogova, A. V.; Ataullakhanov, F. I.; Gudimchuk, N. B.; Vorobyev, Ivan.

In: Molecular biology of the cell, Vol. 30, No. 17, 01.08.2019, p. 2105-2114.

Research output: Contribution to journalArticle

Mustyatsa, VV, Kostarev, AV, Tvorogova, AV, Ataullakhanov, FI, Gudimchuk, NB & Vorobyev, I 2019, 'Fine structure and dynamics of EB3 binding zones on microtubules in fibroblast cells', Molecular biology of the cell, vol. 30, no. 17, pp. 2105-2114. https://doi.org/10.1091/mbc.E18-11-0723
Mustyatsa, V. V. ; Kostarev, A. V. ; Tvorogova, A. V. ; Ataullakhanov, F. I. ; Gudimchuk, N. B. ; Vorobyev, Ivan. / Fine structure and dynamics of EB3 binding zones on microtubules in fibroblast cells. In: Molecular biology of the cell. 2019 ; Vol. 30, No. 17. pp. 2105-2114.
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