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. 1988 Jan 1;106(1):141–149. doi: 10.1083/jcb.106.1.141

Enhanced stability of microtubules enriched in detyrosinated tubulin is not a direct function of detyrosination level

PMCID: PMC2114950  PMID: 3276710

Abstract

Interphase cultured monkey kidney (TC-7) cells contain distinct subsets of cellular microtubules (MTs) enriched in posttranslationally detyrosinated (Glu) or tyrosinated (Tyr) alpha tubulin (Gundersen, G. G., M. H. Kalnoski, and J. C. Bulinski. 1984. Cell. 38:779-789). To determine the relative stability of these subsets of MTs, we subjected TC-7 cells to treatments that slowly depolymerized MTs. We found Glu MTs to be more resistant than Tyr MTs to depolymerization by nocodazole in living cells, and to depolymerization by dilution in detergent- permeabilized cell models. However, in cold-treated cells, Glu and Tyr MTs did not differ significantly in their stability. Digestion of permeabilized cell models with pancreatic carboxypeptidase A, to generate Glu MTs from endogenous Tyr MTs, did not significantly alter the resistance of the endogenous Tyr MTs toward dilution-induced depolymerization. Furthermore, in human fibroblasts that contained no distinct Glu MTs, we observed a population of nocodazole-resistant MTs. These data suggest that Glu MTs possess enhanced stability against end- mediated depolymerization, yet detyrosination alone appears to be insufficient to confer this enhanced stability.

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Selected References

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  1. Arce C. A., Hallak M. E., Rodriguez J. A., Barra H. S., Caputto R. Capability of tubulin and microtubules to incorporate and to release tyrosine and phenylalanine and the effect of the incorporation of these amino acids on tubulin assembly. J Neurochem. 1978 Jul;31(1):205–210. doi: 10.1111/j.1471-4159.1978.tb12449.x. [DOI] [PubMed] [Google Scholar]
  2. Argarana C. E., Barra H. S., Caputto R. Tubulinyl-tyrosine carboxypeptidase from chicken brain: properties and partial purification. J Neurochem. 1980 Jan;34(1):114–118. doi: 10.1111/j.1471-4159.1980.tb04628.x. [DOI] [PubMed] [Google Scholar]
  3. Argaraña C. E., Barra H. S., Caputto R. Release of [14C]tyrosine from tubulinyl-[14C]tyrosine by brain extract. Separation of a carboxypeptidase from tubulin-tyrosine ligase. Mol Cell Biochem. 1978 Feb 24;19(1):17–21. doi: 10.1007/BF00231230. [DOI] [PubMed] [Google Scholar]
  4. Barra H. S., Arce C. A., Caputto R. Total tubulin and its aminoacylated and non-aminoacylated forms during the development of rat brain. Eur J Biochem. 1980 Aug;109(2):439–446. doi: 10.1111/j.1432-1033.1980.tb04813.x. [DOI] [PubMed] [Google Scholar]
  5. Barra H. S., Rodriguez J. A., Arce C. A., Caputto R. A soluble preparation from rat brain that incorporates into its own proteins ( 14 C)arginine by a ribonuclease-sensitive system and ( 14 C)tyrosine by a ribonuclease-insensitive system. J Neurochem. 1973 Jan;20(1):97–108. doi: 10.1111/j.1471-4159.1973.tb12108.x. [DOI] [PubMed] [Google Scholar]
  6. Bulinski J. C., Borisy G. G. Widespread distribution of a 210,000 mol wt microtubule-associated protein in cells and tissues of primates. J Cell Biol. 1980 Dec;87(3 Pt 1):802–808. doi: 10.1083/jcb.87.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carlier M. F., Pantaloni D. Kinetic analysis of guanosine 5'-triphosphate hydrolysis associated with tubulin polymerization. Biochemistry. 1981 Mar 31;20(7):1918–1924. doi: 10.1021/bi00510a030. [DOI] [PubMed] [Google Scholar]
  8. Cassimeris L. U., Wadsworth P., Salmon E. D. Dynamics of microtubule depolymerization in monocytes. J Cell Biol. 1986 Jun;102(6):2023–2032. doi: 10.1083/jcb.102.6.2023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deanin G. G., Thompson W. C., Gordon M. W. Tyrosyltubulin ligase activity in brain, skeletal muscle, and liver of the developing chick. Dev Biol. 1977 May;57(1):230–233. doi: 10.1016/0012-1606(77)90370-0. [DOI] [PubMed] [Google Scholar]
  10. Dentler W. L. Attachment of the cap to the central microtubules of Tetrahymena cilia. J Cell Sci. 1984 Mar;66:167–173. doi: 10.1242/jcs.66.1.167. [DOI] [PubMed] [Google Scholar]
  11. Flavin M., Murofushi H. Tyrosine incorporation in tubulin. Methods Enzymol. 1984;106:223–237. doi: 10.1016/0076-6879(84)06024-9. [DOI] [PubMed] [Google Scholar]
  12. Forrest G. L., Klevecz R. R. Tyrosyltubulin ligase and colchicine binding activity in synchronized Chinese hamster cells. J Cell Biol. 1978 Aug;78(2):441–450. doi: 10.1083/jcb.78.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geuens G., Gundersen G. G., Nuydens R., Cornelissen F., Bulinski J. C., DeBrabander M. Ultrastructural colocalization of tyrosinated and detyrosinated alpha-tubulin in interphase and mitotic cells. J Cell Biol. 1986 Nov;103(5):1883–1893. doi: 10.1083/jcb.103.5.1883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gundersen G. G., Bulinski J. C. Distribution of tyrosinated and nontyrosinated alpha-tubulin during mitosis. J Cell Biol. 1986 Mar;102(3):1118–1126. doi: 10.1083/jcb.102.3.1118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gundersen G. G., Bulinski J. C. Microtubule arrays in differentiated cells contain elevated levels of a post-translationally modified form of tubulin. Eur J Cell Biol. 1986 Dec;42(2):288–294. [PubMed] [Google Scholar]
  16. Gundersen G. G., Kalnoski M. H., Bulinski J. C. Distinct populations of microtubules: tyrosinated and nontyrosinated alpha tubulin are distributed differently in vivo. Cell. 1984 Oct;38(3):779–789. doi: 10.1016/0092-8674(84)90273-3. [DOI] [PubMed] [Google Scholar]
  17. Gundersen G. G., Khawaja S., Bulinski J. C. Postpolymerization detyrosination of alpha-tubulin: a mechanism for subcellular differentiation of microtubules. J Cell Biol. 1987 Jul;105(1):251–264. doi: 10.1083/jcb.105.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hill T. L., Carlier M. F. Steady-state theory of the interference of GTP hydrolysis in the mechanism of microtubule assembly. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7234–7238. doi: 10.1073/pnas.80.23.7234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kilmartin J. V., Wright B., Milstein C. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol. 1982 Jun;93(3):576–582. doi: 10.1083/jcb.93.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kumar N., Flavin M. Modulation of some parameters of assembly of microtubules in vitro by tyrosinolation of tubulin. Eur J Biochem. 1982 Nov;128(1):215–222. doi: 10.1111/j.1432-1033.1982.tb06954.x. [DOI] [PubMed] [Google Scholar]
  21. Kumar N., Flavin M. Preferential action of a brain detyrosinolating carboxypeptidase on polymerized tubulin. J Biol Chem. 1981 Jul 25;256(14):7678–7686. [PubMed] [Google Scholar]
  22. LeDizet M., Piperno G. Cytoplasmic microtubules containing acetylated alpha-tubulin in Chlamydomonas reinhardtii: spatial arrangement and properties. J Cell Biol. 1986 Jul;103(1):13–22. doi: 10.1083/jcb.103.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mandelkow E. M., Mandelkow E. Unstained microtubules studied by cryo-electron microscopy. Substructure, supertwist and disassembly. J Mol Biol. 1985 Jan 5;181(1):123–135. doi: 10.1016/0022-2836(85)90330-4. [DOI] [PubMed] [Google Scholar]
  24. Mitchison T. J., Kirschner M. W. Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation. J Cell Biol. 1985 Sep;101(3):766–777. doi: 10.1083/jcb.101.3.766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mitchison T., Kirschner M. Dynamic instability of microtubule growth. Nature. 1984 Nov 15;312(5991):237–242. doi: 10.1038/312237a0. [DOI] [PubMed] [Google Scholar]
  26. Moyer S. A., Baker S. C., Lessard J. L. Tubulin: a factor necessary for the synthesis of both Sendai virus and vesicular stomatitis virus RNAs. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5405–5409. doi: 10.1073/pnas.83.15.5405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nath J., Flavin M., Gallin J. I. Tubulin tyrosinolation in human polymorphonuclear leukocytes: studies in normal subjects and in patients with the Chediak-Higashi syndrome. J Cell Biol. 1982 Nov;95(2 Pt 1):519–526. doi: 10.1083/jcb.95.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nath J., Flavin M. Tubulin tyrosylation in vivo and changes accompanying differentiation of cultured neuroblastoma-glioma hybrid cells. J Biol Chem. 1979 Nov 25;254(22):11505–11510. [PubMed] [Google Scholar]
  29. Piperno G., LeDizet M., Chang X. J. Microtubules containing acetylated alpha-tubulin in mammalian cells in culture. J Cell Biol. 1987 Feb;104(2):289–302. doi: 10.1083/jcb.104.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raybin D., Flavin M. Enzyme which specifically adds tyrosine to the alpha chain of tubulin. Biochemistry. 1977 May 17;16(10):2189–2194. doi: 10.1021/bi00629a023. [DOI] [PubMed] [Google Scholar]
  31. Raybin D., Flavin M. Modification of tubulin by tyrosylation in cells and extracts and its effect on assembly in vitro. J Cell Biol. 1977 May;73(2):492–504. doi: 10.1083/jcb.73.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rodriguez J. A., Borisy G. G. Modification of the C-terminus of brain tubulin during development. Biochem Biophys Res Commun. 1978 Jul 28;83(2):579–586. doi: 10.1016/0006-291x(78)91029-x. [DOI] [PubMed] [Google Scholar]
  33. Rodriguez J. A., Borisy G. G. Tyrosination state of free tubulin subunits and tubulin disassembled from microtubules of rat brain tissue. Biochem Biophys Res Commun. 1979 Aug 13;89(3):893–899. doi: 10.1016/0006-291x(79)91862-x. [DOI] [PubMed] [Google Scholar]
  34. Schulze E., Kirschner M. Dynamic and stable populations of microtubules in cells. J Cell Biol. 1987 Feb;104(2):277–288. doi: 10.1083/jcb.104.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schulze E., Kirschner M. Microtubule dynamics in interphase cells. J Cell Biol. 1986 Mar;102(3):1020–1031. doi: 10.1083/jcb.102.3.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Soltys B. J., Borisy G. G. Polymerization of tubulin in vivo: direct evidence for assembly onto microtubule ends and from centrosomes. J Cell Biol. 1985 May;100(5):1682–1689. doi: 10.1083/jcb.100.5.1682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Thompson W. C., Asai D. J., Carney D. H. Heterogeneity among microtubules of the cytoplasmic microtubule complex detected by a monoclonal antibody to alpha tubulin. J Cell Biol. 1984 Mar;98(3):1017–1025. doi: 10.1083/jcb.98.3.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Webster D. R., Gundersen G. G., Bulinski J. C., Borisy G. G. Assembly and turnover of detyrosinated tubulin in vivo. J Cell Biol. 1987 Jul;105(1):265–276. doi: 10.1083/jcb.105.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wehland J., Willingham M. C., Sandoval I. V. A rat monoclonal antibody reacting specifically with the tyrosylated form of alpha-tubulin. I. Biochemical characterization, effects on microtubule polymerization in vitro, and microtubule polymerization and organization in vivo. J Cell Biol. 1983 Nov;97(5 Pt 1):1467–1475. doi: 10.1083/jcb.97.5.1467. [DOI] [PMC free article] [PubMed] [Google Scholar]

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