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. 1994 Dec 1;127(5):1289–1299. doi: 10.1083/jcb.127.5.1289

Effect on microtubule dynamics of XMAP230, a microtubule-associated protein present in Xenopus laevis eggs and dividing cells [published erratum appears in J Cell Biol 1995 Mar;128(5):following 988]

PMCID: PMC2120251  PMID: 7962090

Abstract

The reorganization from a radial [corrected] interphase microtubule (MT) network into a bipolar spindle at the onset of mitosis involves a dramatic change in MT dynamics. Microtubule-associated proteins (MAPs) and other factors are thought to regulate MT dynamics both in interphase and in mitosis. In this study we report the purification and functional in vitro characterization of a 230-KD MAP from Xenopus egg extract (XMAP230). This protein is present in eggs, oocytes, testis and a Xenopus tissue culture cell line. It is apparently absent from non- dividing cells in which an immunologically related 200-kD protein is found. XMAP230 is composed of two isoforms with slightly different molecular masses and pIs. It is localized to interphase MTs, dissociates from MTs at the onset of prophase and specifically binds to spindle MTs during metaphase and anaphase. The dissociation constant of XMAP230 is 500 nM, the stoichiometry of binding to MTs is between 1:8 and 1:4, and the in vivo concentration is approximately 200 nM. Both isoforms are phosphorylated and have reduced affinity for microtubules in mitotic extracts. Analysis of the effect of XMAP230 on MT dynamics by video microscopy shows that it increases the growth rate, decreases the shrinking rate of MTs and strongly suppresses catastrophes. These results suggest that in vivo, XMAP230 participates in the control of the MT elongation rate, stabilizes MTs and locally modulates MT dynamics during mitosis.

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Selected References

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  1. Allen R. D., Allen N. S., Travis J. L. Video-enhanced contrast, differential interference contrast (AVEC-DIC) microscopy: a new method capable of analyzing microtubule-related motility in the reticulopodial network of Allogromia laticollaris. Cell Motil. 1981;1(3):291–302. doi: 10.1002/cm.970010303. [DOI] [PubMed] [Google Scholar]
  2. Belmont L. D., Hyman A. A., Sawin K. E., Mitchison T. J. Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts. Cell. 1990 Aug 10;62(3):579–589. doi: 10.1016/0092-8674(90)90022-7. [DOI] [PubMed] [Google Scholar]
  3. Bloom G. S., Luca F. C., Vallee R. B. Identification of high molecular weight microtubule-associated proteins in anterior pituitary tissue and cells using taxol-dependent purification combined with microtubule-associated protein specific antibodies. Biochemistry. 1985 Jul 16;24(15):4185–4191. doi: 10.1021/bi00336a055. [DOI] [PubMed] [Google Scholar]
  4. Bornens M., Paintrand M., Berges J., Marty M. C., Karsenti E. Structural and chemical characterization of isolated centrosomes. Cell Motil Cytoskeleton. 1987;8(3):238–249. doi: 10.1002/cm.970080305. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Bré M. H., Karsenti E. Effects of brain microtubule-associated proteins on microtubule dynamics and the nucleating activity of centrosomes. Cell Motil Cytoskeleton. 1990;15(2):88–98. doi: 10.1002/cm.970150205. [DOI] [PubMed] [Google Scholar]
  7. Bré M. H., Kreis T. E., Karsenti E. Control of microtubule nucleation and stability in Madin-Darby canine kidney cells: the occurrence of noncentrosomal, stable detyrosinated microtubules. J Cell Biol. 1987 Sep;105(3):1283–1296. doi: 10.1083/jcb.105.3.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Butner K. A., Kirschner M. W. Tau protein binds to microtubules through a flexible array of distributed weak sites. J Cell Biol. 1991 Nov;115(3):717–730. doi: 10.1083/jcb.115.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Domínguez J. E., Buendia B., López-Otín C., Antony C., Karsenti E., Avila J. A protein related to brain microtubule-associated protein MAP1B is a component of the mammalian centrosome. J Cell Sci. 1994 Feb;107(Pt 2):601–611. [PubMed] [Google Scholar]
  10. Drechsel D. N., Hyman A. A., Cobb M. H., Kirschner M. W. Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau. Mol Biol Cell. 1992 Oct;3(10):1141–1154. doi: 10.1091/mbc.3.10.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Evans L., Mitchison T., Kirschner M. Influence of the centrosome on the structure of nucleated microtubules. J Cell Biol. 1985 Apr;100(4):1185–1191. doi: 10.1083/jcb.100.4.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Faruki S., Karsenti E. Purification of microtubule proteins from Xenopus egg extracts: identification of a 230K MAP4-like protein. Cell Motil Cytoskeleton. 1994;28(2):108–118. doi: 10.1002/cm.970280203. [DOI] [PubMed] [Google Scholar]
  13. Fernandez A., Brautigan D. L., Lamb N. J. Protein phosphatase type 1 in mammalian cell mitosis: chromosomal localization and involvement in mitotic exit. J Cell Biol. 1992 Mar;116(6):1421–1430. doi: 10.1083/jcb.116.6.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gard D. L., Kirschner M. W. A microtubule-associated protein from Xenopus eggs that specifically promotes assembly at the plus-end. J Cell Biol. 1987 Nov;105(5):2203–2215. doi: 10.1083/jcb.105.5.2203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gliksman N. R., Parsons S. F., Salmon E. D. Okadaic acid induces interphase to mitotic-like microtubule dynamic instability by inactivating rescue. J Cell Biol. 1992 Dec;119(5):1271–1276. doi: 10.1083/jcb.119.5.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hirokawa N., Shiomura Y., Okabe S. Tau proteins: the molecular structure and mode of binding on microtubules. J Cell Biol. 1988 Oct;107(4):1449–1459. doi: 10.1083/jcb.107.4.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Izant J. G., Weatherbee J. A., McIntosh J. R. A microtubule-associated protein antigen unique to mitotic spindle microtubules in PtK1 cells. J Cell Biol. 1983 Feb;96(2):424–434. doi: 10.1083/jcb.96.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kirschner M., Mitchison T. Beyond self-assembly: from microtubules to morphogenesis. Cell. 1986 May 9;45(3):329–342. doi: 10.1016/0092-8674(86)90318-1. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Mastronarde D. N., McDonald K. L., Ding R., McIntosh J. R. Interpolar spindle microtubules in PTK cells. J Cell Biol. 1993 Dec;123(6 Pt 1):1475–1489. doi: 10.1083/jcb.123.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miller L., Daniel J. C. Comparison of in vivo and in vitro ribosomal RNA synthesis in nucleolar mutants of Xenopus laevis. In Vitro. 1977 Sep;13(9):557–563. doi: 10.1007/BF02627851. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Mitchison T., Kirschner M. Microtubule assembly nucleated by isolated centrosomes. Nature. 1984 Nov 15;312(5991):232–237. doi: 10.1038/312232a0. [DOI] [PubMed] [Google Scholar]
  24. Murofushi H., Kotani S., Aizawa H., Hisanaga S., Hirokawa N., Sakai H. Purification and characterization of a 190-kD microtubule-associated protein from bovine adrenal cortex. J Cell Biol. 1986 Nov;103(5):1911–1919. doi: 10.1083/jcb.103.5.1911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murray A. W. Cell cycle extracts. Methods Cell Biol. 1991;36:581–605. [PubMed] [Google Scholar]
  26. Neant I., Guerrier P. 6-Dimethylaminopurine blocks starfish oocyte maturation by inhibiting a relevant protein kinase activity. Exp Cell Res. 1988 May;176(1):68–79. doi: 10.1016/0014-4827(88)90121-8. [DOI] [PubMed] [Google Scholar]
  27. Parysek L. M., Wolosewick J. J., Olmsted J. B. MAP 4: a microtubule-associated protein specific for a subset of tissue microtubules. J Cell Biol. 1984 Dec;99(6):2287–2296. doi: 10.1083/jcb.99.6.2287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pryer N. K., Walker R. A., Skeen V. P., Bourns B. D., Soboeiro M. F., Salmon E. D. Brain microtubule-associated proteins modulate microtubule dynamic instability in vitro. Real-time observations using video microscopy. J Cell Sci. 1992 Dec;103(Pt 4):965–976. doi: 10.1242/jcs.103.4.965. [DOI] [PubMed] [Google Scholar]
  29. Shiina N., Moriguchi T., Ohta K., Gotoh Y., Nishida E. Regulation of a major microtubule-associated protein by MPF and MAP kinase. EMBO J. 1992 Nov;11(11):3977–3984. doi: 10.1002/j.1460-2075.1992.tb05491.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ulloa L., Avila J., Díaz-Nido J. Heterogeneity in the phosphorylation of microtubule-associated protein MAP1B during rat brain development. J Neurochem. 1993 Sep;61(3):961–972. doi: 10.1111/j.1471-4159.1993.tb03609.x. [DOI] [PubMed] [Google Scholar]
  31. Ulloa L., Díaz-Nido J., Avila J. Depletion of casein kinase II by antisense oligonucleotide prevents neuritogenesis in neuroblastoma cells. EMBO J. 1993 Apr;12(4):1633–1640. doi: 10.1002/j.1460-2075.1993.tb05808.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vale R. D. Severing of stable microtubules by a mitotically activated protein in Xenopus egg extracts. Cell. 1991 Feb 22;64(4):827–839. doi: 10.1016/0092-8674(91)90511-v. [DOI] [PubMed] [Google Scholar]
  33. Verde F., Dogterom M., Stelzer E., Karsenti E., Leibler S. Control of microtubule dynamics and length by cyclin A- and cyclin B-dependent kinases in Xenopus egg extracts. J Cell Biol. 1992 Sep;118(5):1097–1108. doi: 10.1083/jcb.118.5.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Verde F., Labbé J. C., Dorée M., Karsenti E. Regulation of microtubule dynamics by cdc2 protein kinase in cell-free extracts of Xenopus eggs. Nature. 1990 Jan 18;343(6255):233–238. doi: 10.1038/343233a0. [DOI] [PubMed] [Google Scholar]
  35. Walker R. A., O'Brien E. T., Pryer N. K., Soboeiro M. F., Voter W. A., Erickson H. P., Salmon E. D. Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies. J Cell Biol. 1988 Oct;107(4):1437–1448. doi: 10.1083/jcb.107.4.1437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Walker R. A., Pryer N. K., Salmon E. D. Dilution of individual microtubules observed in real time in vitro: evidence that cap size is small and independent of elongation rate. J Cell Biol. 1991 Jul;114(1):73–81. doi: 10.1083/jcb.114.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]

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