Advertisement
Journal Article
Detyrosinated (Glu) microtubules are stabilized by an ATP-sensitive plus-end cap
A.S. Infante, M.S. Stein, Y. Zhai, G.G. Borisy, G.G. Gundersen
Journal of Cell Science 2000 113: 3907-3919;

Summary

Many cell types contain a subset of long-lived, ‘stable’ microtubules that differ from dynamic microtubules in that they are enriched in post-translationally detyrosinated tubulin (Glu-tubulin). Elevated Glu tubulin does not stabilize the microtubules and the mechanism for the stability of Glu microtubules is not known. We used detergent-extracted cell models to investigate the nature of Glu microtubule stability. In these cell models, Glu microtubules did not incorporate exogenously added tubulin subunits on their distal ends, while >70% of the bulk microtubules did. Ca(2+)-generated fragments of Glu microtubules incorporated tubulin, showing that Glu microtubule ends are capped. Consistent with this, Glu microtubules in cell models were resistant to dilution-induced breakdown. Known microtubule end-associated proteins (EB1, APC, p150(Glued) and vinculin focal adhesions) were not localized on Glu microtubule ends. ATP, but not nonhydrolyzable analogues, induced depolymerization of Glu microtubules in cell models. Timelapse and photobleaching studies showed that ATP triggered subunit loss from the plus end. ATP breakdown of Glu microtubules was inhibited by AMP-PNP and vanadate, but not by kinase or other inhibitors. Additional experiments showed that conventional kinesin or kif3 were not involved in Glu microtubule capping. We conclude that Glu microtubules are stabilized by a plus-end cap that includes an ATPase with properties similar to kinesins.

REFERENCES

    1. Banerjee, A.,
    2. Roach, M. C.,
    3. Trcka, P. and
    4. Ludueña, R. F.
    ) (1990). Increased microtubule assembly in bovine brain tubulin lacking the type III isotype of-tubulin. J. Biol. Chem 265, 1794–1799
    OpenUrlAbstract/FREE Full Text
    1. Bershadsky, A. D. and
    2. Gelfand, V. I.
    (1981). ATP-dependent regulation of cytoplasmic microtubule disassembly. Proc. Natl. Acad. Sci. USA 78, 3610–3613
    OpenUrlAbstract/FREE Full Text
    1. Berrueta, L.,
    2. Kraeft, S.-T.,
    3. Tirnauer, J. S.,
    4. Schuyler, S. C.,
    5. Chen, L. B.,
    6. Hill, D. E.,
    7. Pellman, D. and
    8. Bierer, B. E.
    (1998). The adenomatous polyposis coli-binding protein EB1 is associated with cytoplasmic and spindle microtubules. Proc. Natl. Acad. Sci. USA 95, 10596–10601
    OpenUrlAbstract/FREE Full Text
    1. Brinkley, B. R. and
    2. Cartwright, J.
    (1975). Cold-labile and cold-stable microtubules in the mitotic spindle of mammalian cells. Ann. NY Acad. Sci 253, 428–439
    OpenUrlCrossRefPubMedWeb of Science
    1. Bulinski, J. C. and
    2. Gundersen, G. G.
    (1991). Stabilization and post-translational modification of microtubules during cellular morphogenesis. BioEssays 13, 285–293
    OpenUrlCrossRefPubMedWeb of Science
    1. Chapin, S. J. and
    2. Bulinski, J. C.
    (1994). Cellular microtubules heterogenous in their content of MAP4. Cell Motil. Cytoskel 27, 133–149
    OpenUrlCrossRefPubMedWeb of Science
    1. Cook, T. A.,
    2. Nagasaki, T. and
    3. Gundersen, G. G.
    (1998). Rho guanosine triphosphate mediates the selective stabilization of microtubules induced by lysophosphatidic acid. J. Cell Biol 141, 175–185
    OpenUrlAbstract/FREE Full Text
    1. Desai, A.,
    2. Verma, S.,
    3. Mitchison, T. J. and
    4. Walczak, C. E.
    (1999). Kin I kinesins are microtubule-destabilizing enzymes. Cell 96, 69–78
    OpenUrlCrossRefPubMedWeb of Science
    1. DeZwann, T. M.,
    2. Ellingson, E.,
    3. Pellman, D. and
    4. Roof, D. M.
    (1997). Kinesin related KIP3 of Saccharomyces cerevisiae is required for a distinct step in nuclear migration. J. Cell Biol 138, 1023–1040
    OpenUrlAbstract/FREE Full Text
    1. Gorbsky, G. J.,
    2. Sammak, P. J. and
    3. Borisy, G. G.
    (1987). Chromosomes move poleward in anaphase along stationary microtubules that coordinately disassemble from their kinetochore ends. J. Cell Biol 104, 9–18
    OpenUrlAbstract/FREE Full Text
    1. Gundersen, G. G.,
    2. Kalnoski, M. H. and
    3. Bulinski, J. C.
    (1984). Distinct populations of microtubules: Tyrosinated and nontyrosinated alpha tubulin are distributed differently in vivo. Cell 38, 779–789
    OpenUrlCrossRefPubMedWeb of Science
    1. Gundersen, G. G. and
    2. Bulinski., J. C.
    (1986). Microtubule arrays indifferentiated cells contain elevated levels of a post-translationally modified form of tubulin. Eur. J. Biol 42, 288–294
    OpenUrlPubMedWeb of Science
    1. Gundersen, G. G.,
    2. Khawaja, S. and
    3. Bulinski, J. C.
    (1987). Postpolymerization detyrosination of alpha-tubulin: A mechanism for subcellular differentiation of microtubules. J. Cell Biol 105, 251–264
    OpenUrlAbstract/FREE Full Text
    1. Gurland, G. and
    2. Gundersen, G. G.
    (1993). Protein phosphatase inhibitors induce the selective breakdown of stable MTs in fibroblasts and epithelial cells. Proc. Natl. Acad. Sci. USA 90, 8827–8831
    OpenUrlAbstract/FREE Full Text
    1. Gurland, G. and
    2. Gundersen, G. G.
    (1995). Stable, detyrosinated microtubules function to localize vimentin intermediate filaments in fibroblasts. J. Cell Biol 131, 1275–1290
    OpenUrlAbstract/FREE Full Text
    1. Heil-Chapdelaine, R. A.,
    2. Adames, N. R. and
    3. Cooper, J. A.
    (1999). Formin' the connection between microtubules and the cell cortex. J. Cell Biol 144, 809–811
    OpenUrlFREE Full Text
    1. Hirokawa, N.
    (1994). Microtubule organization and dynamics dependent on microtubule-associated proteins. Curr. Opin. Cell Biol 6, 74–81
    OpenUrlCrossRefPubMedWeb of Science
    1. Ingold, A. L.,
    2. Cohn, S. A. and
    3. Scholey, J. M.
    (1988). Inhibition of kinesin-driven microtubule motility by monoclonal antibodies to kinesin heavy chains. J. Cell Biol 107, 2657–2667
    OpenUrlAbstract/FREE Full Text
    1. Kaverina, I.,
    2. Rottner, K. and
    3. Small, J. V.
    (1998). Targeting, capture and stabilization of microtubules at early focal adhesions. J. Cell Biol 142, 181–190
    OpenUrlAbstract/FREE Full Text
    1. Khawaja, S.,
    2. Gundersen, G. G. and
    3. Bulinski, J. C.
    (1988). Enhanced stability of microtubules enriched in detyrosinated tubulin is not a direct function of detyrosination level. J. Cell Biol 106, 141–149
    OpenUrlAbstract/FREE Full Text
    1. Kilmartin, J. V.,
    2. Wright, B. and
    3. Milstein, C.
    (1982). Rat monoclonal anti-tubulin antibodies derived by using a new nonsecreting rat cell line. J. Cell Biol 93, 576–582
    OpenUrlAbstract/FREE Full Text
    1. Kreis, T. E.
    (1987). Microtubules containing detyrosinated tubulin are less dynamic. EMBO J 6, 2597–2606
    OpenUrlPubMedWeb of Science
    1. Korinek, W. S.,
    2. Copeland, M. J.,
    3. Chadhuri, A. and
    4. Chant, J.
    (2000). Molecular linkage underlying microtubule orientation toward cortical sites. Science 287, 2257–2259
    OpenUrlAbstract/FREE Full Text
    1. Lee, L.,
    2. Klee, S. K.,
    3. Evangelista, M.,
    4. Boone, C. and
    5. Pellman, D.
    (1999). Control of mitotic spindle position by the Saccharomyces cerevisiae formin Bni1p. J. Cell Biol 144, 947–961
    OpenUrlAbstract/FREE Full Text
    1. Lee, L.,
    2. Tirnauer, J. S.,
    3. Li, J.,
    4. Schuyler, S. C.,
    5. Liu, J. Y. and
    6. Pellman, D.
    (2000). Positioning of the mitotic spindle by cortical-microtubule capture mechanism. Science 287, 2260–2262
    OpenUrlAbstract/FREE Full Text
    1. Liao, G.,
    2. Kreitzer, G.,
    3. Cook, T. A. and
    4. Gundersen, G. G.
    (1999). A signal transduction pathway involved in microtubule-mediated cell polarization. FASEB J 13, 257–.
    OpenUrl
    1. Lombillo, V. A.,
    2. Stewart, R. J. and
    3. McIntosh, J. R.
    (1995). Minus-end-directed motion of kinesin-coated microspheres driven by microtubule depolymerization. Nature 373, 161–164
    OpenUrlCrossRefPubMedWeb of Science
    1. Margolis, R. L.,
    2. Rauch, C. T. and
    3. Job, D.
    (1986). Purification and assay of a 145-kDa protein (STOP145) with microtubule-stabilizing and motility behavior. Proc. Natl. Acad. Sci. USA 83, 639–643
    OpenUrlAbstract/FREE Full Text
    1. McIntosh, J. R. and
    2. Porter, M. E.
    (1989). Enzymes for microtubule-dependent motility. J. Biol. Chem 264, 6001–6004
    OpenUrlFREE Full Text
    1. Merrick, S. E.,
    2. Trojanowski, J. Q. and
    3. Lee, V. M.-Y.
    (1997). Selective destruction of stable microtubules and axons by inhibitors of protein serine/threonine phosphatases in cultured human neurons (NT2N cells). J. Neurosci 17, 5726–5737
    OpenUrlAbstract/FREE Full Text
    1. Mikhailov, A. and
    2. Gundersen, G. G.
    (1995). Centripital transport of microtubules in motile cells. Cell Motil. Cytoskel 32, 173–186
    OpenUrlCrossRefPubMedWeb of Science
    1. Mikhailov, A. and
    2. Gundersen, G. G.
    (1998). Direct imaging of microtubule behavior in locomoting fibroblasts treated with nocodazole or taxol reveals a relationship between microtubule dynamics and lamellipodia formation. Cell Motil. Cytoskel 41, 325–340
    OpenUrlCrossRefPubMedWeb of Science
    1. Miller, J. M.,
    2. Wang, W.,
    3. Balczon, R. and
    4. Dentler, W. L.
    (1990). Ciliary microtubule capping structures contain a mammalian kinetochore antigen. J. Cell Biol 110, 703–714
    OpenUrlAbstract/FREE Full Text
    1. Mitchison, T. J.
    (1988). Microtubule dynamics and kinetochore function in mitosis. Ann. Rev. Cell Biol 4, 527–549
    OpenUrlCrossRefWeb of Science
    1. Mitchison, T. J. and
    2. Salmon, E. D.
    (1992). Poleward kinetochore fiber movement occurs during both metaphase and anaphase-A in newt lung cell mitosis. J. Cell Biol 119, 569–82
    OpenUrlAbstract/FREE Full Text
    1. Morrison, E. E.,
    2. Wardleworth, B. N.,
    3. Askham, J. M.,
    4. Markham, A. F. and
    5. Meredith, M. E.
    (1998). EB1, a protein which interacts with APC tumor suppressor, is associated with the microtubule cytoskeletons throughout the cell cycle. Oncogene 17, 3471–3477
    OpenUrlCrossRefPubMedWeb of Science
    1. Nathke, I. S.,
    2. Adams, C. L.,
    3. Polakis, P.,
    4. Sellin, J. H. and
    5. Nelson, W. J.
    (1996). The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration. J. Cell Biol 134, 165–179
    OpenUrlAbstract/FREE Full Text
    1. Perez, F.,
    2. Diamantopoulis, G. S.,
    3. Stadler, R. and
    4. Kreis, T. E.
    (1999). CLIP-170 highlights growing microtubule ends in vivo. Cell 96, 517–527
    OpenUrlCrossRefPubMedWeb of Science
    1. Pfarr, C. M.,
    2. Coue, M.,
    3. Grissom, R. M.,
    4. Hays, T. S.,
    5. Porter, M. E. and
    6. McIntosh, J. R.
    (1990). Cytoplasmic dynein is localized to kinetochores during mitosis. Nature 345, 263–265
    OpenUrlCrossRefPubMed
    1. Rodionov, V. I.,
    2. Lim, S.-S.,
    3. Gelfand, V. and
    4. Borisy, G. G.
    (1994). Microtubule dynamics in fish melanophores. J. Cell Biol 126, 1455–1464
    OpenUrlAbstract/FREE Full Text
    1. Schliwa, M.,
    2. Euteneuer, U.,
    3. Bulinski, J. C. and
    4. Izant, J. G.
    (1981). Calcium lability of cytoplasmic microtubules and its modulation by microtubule-associated proteins. Proc. Natl. Acad. Sci. USA 78, 1037–1041
    OpenUrlAbstract/FREE Full Text
    1. Schulze, E. and
    2. Kirschner, M.
    (1987). Dynamic and stable populations of microtubules in cells. J. Cell Biol 104, 277–288
    OpenUrlAbstract/FREE Full Text
    1. Schulze, E.,
    2. Asai, D.,
    3. Bulinski, J. C. and
    4. Kirschner, M.
    (1987). Post-translational modifications and microtubule stability. J. Cell Biol 105, 2167–2178
    OpenUrlAbstract/FREE Full Text
    1. Shelden, E. and
    2. Wadsworth, P.
    (1993). Observation and quantification of individual microtubule behavior in vivo are cell-type specific. J. Cell Biol 120, 935–945
    OpenUrlAbstract/FREE Full Text
    1. Shiff, P. B. and
    2. Horowitz, S. B.
    (1980). Taxol stabilizes microtubules in mouse fibroblast cells. Proc. Natl. Acad. Sci. USA 77, 1561–1565
    OpenUrlAbstract/FREE Full Text
    1. Skoufias, D. A. and
    2. Wilson, L.
    (1998). Assembly and colchicine binding characteristics of tubulin with maximally tyrosinated and detyrosinated alpha-tubulins. Arch. Biochem. Biophys 351, 115–122
    OpenUrlCrossRefPubMedWeb of Science
    1. Steuer, E. R.,
    2. Wordeman, L.,
    3. Shroer, T. A. and
    4. Sheetz, M. P.
    (1990). Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature 345, 266–268
    OpenUrlCrossRefPubMed
    1. Thompson, W. C.,
    2. Asai, D. J. and
    3. Carney, D. H.
    (1984). Heterogeneity among microtubules of the cytoplasmic microtubule complex detected with a monoclonal antibody to alpha tubulin. J. Cell Biol 98, 1017–1025
    OpenUrlAbstract/FREE Full Text
    1. Vaughan, K. T.,
    2. Tynan, S. H.,
    3. Faulkner, N. E.,
    4. Echeverri, C. J. and
    5. Vallee, R. B.
    (1999). Colocalization of cytoplasmic dynein with dynactin and CLIP-170 at microtubule distal ends. J. Cell Sci 112, 1437–1447
    OpenUrlAbstract/FREE Full Text
    1. Wang, W.,
    2. Himes, R. H. and
    3. Dentler, W. L.
    (1994). The binding of a ciliary microtubule plus-end binding protein complex to microtubules is regulated by ciliary protein kinase and phosphatase activities. J. Biol. Chem 269, 21460–21466
    OpenUrlAbstract/FREE Full Text
    1. Webster, D.,
    2. Gundersen, G. G.,
    3. Bulinski, J. C. and
    4. Borisy, G. G.
    (1987). Differential turnover of tyrosinated and detyrosinated microtubules. Proc. Natl. Acad. Sci. USA 84, 9040–9044
    OpenUrlAbstract/FREE Full Text
    1. Webster, D.,
    2. Gundersen, G. G.,
    3. Bulinski, J. C. and
    4. Borisy, G. G.
    (1987). Assembly and turnover of detyrosinated tubulin in vivo. J. Cell Biol 105, 265–276
    OpenUrlAbstract/FREE Full Text
    1. Webster, D. R.,
    2. Wehland, J.,
    3. Weber, K. and
    4. Borisy, G. G.
    (1990). Detyrosination of alpha tubulin does not stabilize microtubules in vivo. J. Cell Biol 111, 113–122
    OpenUrlAbstract/FREE Full Text
    1. Wordeman, L. and
    2. Mitchison, T. J.
    (1995). Identification and partial characterization of mitotic centromere-associated kinesin, a kinesin-related protein that associates with centromeres during mitosis. J. Cell Biol 128, 95–104
    OpenUrlAbstract/FREE Full Text
    1. Yen, T. J.,
    2. Li, G.,
    3. Schaar, B. T.,
    4. Szilak, I. and
    5. Cleveland, D. W.
    (1992). CENP-E is a putative kinetochore motor that accumulates just before mitosis. Nature 359, 536–539
    OpenUrlCrossRefPubMed
Back to top

 Download PDF

Email
Journal Article
Detyrosinated (Glu) microtubules are stabilized by an ATP-sensitive plus-end cap
A.S. Infante, M.S. Stein, Y. Zhai, G.G. Borisy, G.G. Gundersen
Journal of Cell Science 2000 113: 3907-3919;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Follow us on Instagram

Cell science is bursting with beautiful images and over on Instagram, we’re showing them off! Find both JCS and FocalPlane on Instagram for stories and techniques across cell biology.

An interview with Derek Walsh

Professor Derek Walsh is the guest editor of our new special issue Cell Biology of Host-Pathogen Interactions. In an interview, Derek tells us about his work in the field of DNA viruses, the impact of the pandemic on virology and what his role as Guest Editor taught him.

How to improve your scientific writing

"If you are a scientist and you want to succeed, you must become a writer."

How do scientists become master storytellers? We called on our journal Editors, proofreaders and contributors to our community sites for their advice on how to improve your scientific writing.

Meet the preLighters: Jennifer Ann Black

Following the theme of our latest special issue, postdoc Jennifer Ann Black studies replication stress and genome plasticity in Leishmania in Professor Luiz Tosi’s lab in Sao Paolo. We caught up with Jenn (virtually) to hear about her relocation to Brazil mid-pandemic, her research on parasites and what she enjoys about ‘preLighting’.

In our special issue, Chandrakar et al. and Rosazza et al. present their latest work on Leishmania.

Mole – The Corona Files

“There are millions of people around the world who continue to believe that the Terrible Pandemic is a hoax.”

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.

JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.