A polymer-dependent increase in phosphorylation of beta-tubulin accompanies differentiation of a mouse neuroblastoma cell line.

Gard, David L.; Kirschner, Marc W.

doi:10.1083/jcb.100.3.764

Cited by 236 publications

(148 citation statements)

References 41 publications

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“…We demonstrate that the increase in class III β-tubulin isotype is linked to the differentiation process in epithelial cells, as reported for differentiated neuronal cells (Gard and Kirshner, 1985;Joshi and Cleveland, 1989). Little is available on the tubulin isotype composition of normal or tumour cells; moreover, functional differences between isotypes that may be generalized to a variety of cell models have not been identified (Luduena, 1993).…”

Section: supporting

confidence: 52%

Differentiation of human colon cancer cells changes the expression of β-tubulin isotypes and MAPs

et al. 1999

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SummaryThe human colon adenocarcinoma HT29-D4 cell line is an interesting model for studies on epithelial cell differentiation. Undifferentiated cells are malignant proliferating cells, whereas differentiated cells act like epithelial polarized cells. In the present study, we first characterized the action of taxoids on the microtubular network of HT29-D4 cells according to the state of differentiation. Microtubular bundles were found in undifferentiated cells but not in differentiated cells, even with 500-fold higher taxoid concentrations for 96 h. This finding led us to study changes in microtubules according to the polarity status of the cell. E-MAP-115 was expressed only in differentiated cells; expression of β-tubulin isotypes was altered in them relative to undifferentiated cells. Classes I, II, III, IVa and IVb isotypes were expressed in both phenotypes; however, differentiated epithelial cells displayed a specific increase in class III β-tubulin. Thus, the increase in expression of this β-tubulin isotype in differentiated cells is not restricted to neuronal cells. Moreover, these expression changes may reflect a higher stability of microtubular network in differentiated cells, which may explain the lower activity of anti-microtubule agents, independently of the mitotic process. These results indicate that the composition of microtubules should be considered as one of the criteria involved in the response of tumour cells to chemotherapy with anti-microtubule agents.

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Section: supporting

confidence: 52%

Differentiation of human colon cancer cells changes the expression of β-tubulin isotypes and MAPs

et al. 1999

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“…However, tubulin, a highly conserved molecule, contains no significant hydrophobic domains (Cleveland and Sullivan, 1985). Furthermore, known posttranslational modifications of tubulin [e.g., tyrosination (Raybin and Flavin, 1977;Deanin et al, 1980), acetylation (L'Hernault and Rosenbaum, 1985;LeDizet and Piperno, 1991), glutamylation (Edde et al, 1990;Alexander et al, 1991), and phosphorylation (Gard and Kirschner, 1985;Alexander et al, 1991;Peters et al, 1996)] cannot account for membrane association. This has led to the suggestion that the putative membrane tubulin may actually be contaminating cytoplasmic tubulin.…”

Section: Introductionmentioning

confidence: 99%

Posttranslational modification of tubulin by palmitoylation: I. In vivo and cell-free studies.

Caron

1997

MBoC

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It is well established that microtubules interact with intracellular membranes of eukaryotic cells. There is also evidence that tubulin, the major subunit of microtubules, associates directly with membranes. In many cases, this association between tubulin and membranes involves hydrophobic interactions. However, neither primary sequence nor known posttranslational modifications of tubulin can account for such an interaction. The goal of this study was to determine the molecular nature of hydrophobic interactions between tubulin and membranes. Specifically, I sought to identify a posttranslational modification of tubulin that is found in membrane proteins but not in cytoplasmic proteins. One such modification is the covalent attachment of the long chain fatty acid palmitate. The possibility that tubulin is a substrate for palmitoylation was investigated. First, I found that tubulin was palmitoylated in resting platelets and that the level of palmitoylation of tubulin decreased upon activation of platelets with thrombin. Second, to obtain quantities of palmitoylated tubulin required for protein structure analysis, a cell-free system for palmitoylation of tubulin was developed and characterized. The substrates for palmitoylation were nonpolymerized tubulin and tubulin in microtubules assembled with the slowly hydrolyzable GTP analogue guanylyl-(a,3)-methylenediphosphonate. However, tubulin in Taxol-assembled microtubules was not a substrate for palmitoylation. Likewise, palmitoylation of tubulin in the cell-free system was specifically inhibited by the antimicrotubule drugs Colcemid, podophyllotoxin, nocodazole, and vinblastine. These experiments identify a previously unknown posttranslational modification of tubulin that can account for at least one type of hydrophobic interaction with intracellular membranes.

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“…In addition, 1987). The detyrosination, acetylation, and phosphorylation reactions occur primarily on tubulin assembled into MTs (Kumar and Flavin, 1981;L'Hernault and Rosenbaum, 1983;Black et al, 1989;Gard and Kirschner, 1985), while the complementary reactions occur primarily on unassembled dimers (Raybin and Flavin, 1975;L'Hernault and Rosenbaum, 1985;Gard and Kirschner, 1985). Although the restriction of each enzyme activity to one phase of the tubulin assembly cycle suggests a functional relationship between them as well, the relationship between posttranslational modification and MT function (including their increased stability) remains uncertain.…”

Section: Introductionmentioning

confidence: 99%

Detyrosination of alpha tubulin does not stabilize microtubules in vivo [published erratum appears in J Cell Biol 1990 Sep;111(3):1325-6]

Webster

Wehland

Weber

et al. 1990

The Journal of Cell Biology

141

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Abstract.The relationship between alpha tubulin detyrosination and microtubule (MT) stability was examined directly in cultured fibroblasts by experimentally converting the predominantly tyrosinated MT array to a detyrosinated (Glu) array and then assaying MT stability. MTs in mouse Swiss 3T3 cells displayed an increase in Glu immunostaining fluorescence ~1 h after microinjecting antibodies to the tyrosinating enzyme, tubulin tyrosine ligase. Detyrosination progressed to virtual completion after 12 h and persisted for 30-35 h before tyrosinated subunits within MTs were again detected. The stability of these experimentally detyrosinated MTs was tested by first injecting either biotinylated or Xrhodamine-labeled tubulin and then measuring bulk turnover by hapten-mediated immunocytochemistry or fluorescence recovery after photobleaching, respectively. By both methods, turnover was found to be similarly rapid, possessing a half time of ,'~ 3 min. As a final test of MT stability, the level of acetylated tubulin staining in antibody-injected cells was compared with that observed in adjacent, uninjected cells and also with the staining observed in cells whose MTs had been stabilized with taxol. Although intense Glu staining was observed in both injected and taxol-treated cells, increased acetylated tubulin staining was observed only in the taxol-stabilized MTs, indicating that the MTs were not stabilized by detyrosination. Together, these results demonstrated clearly that detyrosination does not directly confer stability on MTs. Therefore, the stable MTs observed in these and other cell lines must have arisen by another mechanism, and may have become posttranslationally modified after their stabilization.

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A polymer-dependent increase in phosphorylation of beta-tubulin accompanies differentiation of a mouse neuroblastoma cell line.

Abstract: We have examined the phosphorylation of cellular microtubule proteins during differentiation and neurite outgrowth in N115 mouse neuroblastoma cells.

Cited by 236 publications

References 41 publications

Differentiation of human colon cancer cells changes the expression of β-tubulin isotypes and MAPs

Differentiation of human colon cancer cells changes the expression of β-tubulin isotypes and MAPs

Posttranslational modification of tubulin by palmitoylation: I. In vivo and cell-free studies.

Detyrosination of alpha tubulin does not stabilize microtubules in vivo [published erratum appears in J Cell Biol 1990 Sep;111(3):1325-6]

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