Crossref journal-article
American Society for Cell Biology (ASCB)
Molecular Biology of the Cell (1076)
Abstract

Posttranslationally modified forms of tubulin accumulate in the subset of stabilized microtubules (MTs) in cells but are not themselves involved in generating MT stability. We showed previously that stabilized, detyrosinated (Glu) MTs function to localize vimentin intermediate filaments (IFs) in fibroblasts. To determine whether tubulin detyrosination or MT stability is the critical element in the preferential association of IFs with Glu MTs, we microinjected nonpolymerizable Glu tubulin into cells. If detyrosination is critical, then soluble Glu tubulin should be a competitive inhibitor of the IF–MT interaction. Before microinjection, Glu tubulin was rendered nonpolymerizable and nontyrosinatable by treatment with iodoacetamide (IAA). Microinjected IAA-Glu tubulin disrupted the interaction of IFs with MTs, as assayed by the collapse of IFs to a perinuclear location, and had no detectable effect on the array of Glu or tyrosinated MTs in cells. Conversely, neither IAA-tyrosinated tubulin nor untreated Glu tubulin, which assembled into MTs, caused collapse of IFs when microinjected. The epitope on Glu tubulin responsible for interfering with the Glu MT–IF interaction was mapped by microinjecting tubulin fragments of α-tubulin. The 14-kDa C-terminal fragment of Glu tubulin (α-C Glu) induced IF collapse, whereas the 36-kDa N-terminal fragment of α-tubulin did not alter the IF array. The epitope required more than the detyrosination site at the C terminus, because a short peptide (a 7-mer) mimicking the C terminus of Glu tubulin did not disrupt the IF distribution. We previously showed that kinesin may mediate the interaction of Glu MTs and IFs. In this study we found that kinesin binding to MTs in vitro was inhibited by the same reagents (i.e., IAA-Glu tubulin and α-C Glu) that disrupted the IF–Glu MT interaction in vivo. These results demonstrate for the first time that tubulin detyrosination functions as a signal for the recruitment of IFs to MTs via a mechanism that is likely to involve kinesin.

Bibliography

Kreitzer, G., Liao, G., & Gundersen, G. G. (1999). Detyrosination of Tubulin Regulates the Interaction of Intermediate Filaments with Microtubules In Vivo via a Kinesin-dependent Mechanism. Molecular Biology of the Cell, 10(4), 1105–1118.

Authors 3
  1. Geri Kreitzer (first)
  2. Guojuan Liao (additional)
  3. Gregg G. Gundersen (additional)
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Dates
Type When
Created 12 years, 2 months ago (July 2, 2013, 7:28 p.m.)
Deposited 3 years, 6 months ago (Feb. 26, 2022, 9:38 p.m.)
Indexed 1 month ago (Aug. 6, 2025, 8:24 a.m.)
Issued 26 years, 5 months ago (April 1, 1999)
Published 26 years, 5 months ago (April 1, 1999)
Published Print 26 years, 5 months ago (April 1, 1999)
Funders 0

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@article{Kreitzer_1999, title={Detyrosination of Tubulin Regulates the Interaction of Intermediate Filaments with Microtubules In Vivo via a Kinesin-dependent Mechanism}, volume={10}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.10.4.1105}, DOI={10.1091/mbc.10.4.1105}, number={4}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Kreitzer, Geri and Liao, Guojuan and Gundersen, Gregg G.}, editor={Kirschner, Marc W.}, year={1999}, month=apr, pages={1105–1118} }