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Centrosome duplication continues in cycloheximide-treated Xenopus blastulae in the absence of a detectable cell cycle.

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Affiliations

  • 1. Department of Biology, University of Utah, Salt Lake City.
      Authors
    • Gard DL 1
    (1 author)

The Journal of Cell Biology, 01 Jun 1990, 110(6):2033-2042
https://doi.org/10.1083/jcb.110.6.2033 PMID: 2190990 PMCID: PMC2116137

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Abstract 

Cycloheximide (500 micrograms/ml) rapidly arrests cleavage, spindle assembly, and cycles of an M-phase-specific histone kinase in early Xenopus blastulae. 2 h after cycloheximide addition, most cells contained two microtubule asters radiating from perinuclear microtubule organizing centers (MTOCs). In contrast, blastomeres treated with cycloheximide for longer periods (3-6 h) contained numerous microtubule asters and MTOCs. Immunofluorescence with an anticentrosome serum and EM demonstrated that the MTOCs in cycloheximide-treated cells were typical centrosomes, containing centrioles and pericentriolar material. We conclude that centrosome duplication continues in cycloheximide-treated Xenopus blastulae in the absence of a detectable cell cycle. In addition, these observations suggest that Xenopus embryos contain sufficient material to assemble 1,000-2,000 centrosomes in the absence of normal protein synthesis.

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J Cell Biol. 1990 Jun 1; 110(6): 2033–2042.
PMCID: PMC2116137
PMID: 2190990

Centrosome duplication continues in cycloheximide-treated Xenopus blastulae in the absence of a detectable cell cycle

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Abstract

Cycloheximide (500 micrograms/ml) rapidly arrests cleavage, spindle assembly, and cycles of an M-phase-specific histone kinase in early Xenopus blastulae. 2 h after cycloheximide addition, most cells contained two microtubule asters radiating from perinuclear microtubule organizing centers (MTOCs). In contrast, blastomeres treated with cycloheximide for longer periods (3-6 h) contained numerous microtubule asters and MTOCs. Immunofluorescence with an anticentrosome serum and EM demonstrated that the MTOCs in cycloheximide-treated cells were typical centrosomes, containing centrioles and pericentriolar material. We conclude that centrosome duplication continues in cycloheximide- treated Xenopus blastulae in the absence of a detectable cell cycle. In addition, these observations suggest that Xenopus embryos contain sufficient material to assemble 1,000-2,000 centrosomes in the absence of normal protein synthesis.

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Selected References

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  • Alvey PL. An investigation of the centriole cycle using 3T3 and CHO cells. J Cell Sci. 1985 Oct;78:147–162. [Abstract] [Google Scholar]
  • Arion D, Meijer L, Brizuela L, Beach D. cdc2 is a component of the M phase-specific histone H1 kinase: evidence for identity with MPF. Cell. 1988 Oct 21;55(2):371–378. [Abstract] [Google Scholar]
  • Brinkley BR. Microtubule organizing centers. Annu Rev Cell Biol. 1985;1:145–172. [Abstract] [Google Scholar]
  • Brinkley BR, Cox SM, Pepper DA, Wible L, Brenner SL, Pardue RL. Tubulin assembly sites and the organization of cytoplasmic microtubules in cultured mammalian cells. J Cell Biol. 1981 Sep;90(3):554–562. [Europe PMC free article] [Abstract] [Google Scholar]
  • Calarco-Gillam PD, Siebert MC, Hubble R, Mitchison T, Kirschner M. Centrosome development in early mouse embryos as defined by an autoantibody against pericentriolar material. Cell. 1983 Dec;35(3 Pt 2):621–629. [Abstract] [Google Scholar]
  • Cicirelli MF, Pelech SL, Krebs EG. Activation of multiple protein kinases during the burst in protein phosphorylation that precedes the first meiotic cell division in Xenopus oocytes. J Biol Chem. 1988 Feb 5;263(4):2009–2019. [Abstract] [Google Scholar]
  • Dabauvalle MC, Doree M, Bravo R, Karsenti E. Role of nuclear material in the early cell cycle of Xenopus embryos. Cell. 1988 Feb 26;52(4):525–533. [Abstract] [Google Scholar]
  • DIRKSEN ER. The presence of centrioles in artificially activated sea urchin eggs. J Biophys Biochem Cytol. 1961 Oct;11:244–247. [Europe PMC free article] [Abstract] [Google Scholar]
  • Dunphy WG, Brizuela L, Beach D, Newport J. The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis. Cell. 1988 Jul 29;54(3):423–431. [Abstract] [Google Scholar]
  • Evans T, Rosenthal ET, Youngblom J, Distel D, Hunt T. Cyclin: a protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division. Cell. 1983 Jun;33(2):389–396. [Abstract] [Google Scholar]
  • Forbes DJ, Kirschner MW, Newport JW. Spontaneous formation of nucleus-like structures around bacteriophage DNA microinjected into Xenopus eggs. Cell. 1983 Aug;34(1):13–23. [Abstract] [Google Scholar]
  • Gard DL, Kirschner MW. A polymer-dependent increase in phosphorylation of beta-tubulin accompanies differentiation of a mouse neuroblastoma cell line. J Cell Biol. 1985 Mar;100(3):764–774. [Europe PMC free article] [Abstract] [Google Scholar]
  • Gautier J, Norbury C, Lohka M, Nurse P, Maller J. Purified maturation-promoting factor contains the product of a Xenopus homolog of the fission yeast cell cycle control gene cdc2+. Cell. 1988 Jul 29;54(3):433–439. [Abstract] [Google Scholar]
  • Gerhart J, Wu M, Kirschner M. Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs. J Cell Biol. 1984 Apr;98(4):1247–1255. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hara K, Tydeman P, Kirschner M. A cytoplasmic clock with the same period as the division cycle in Xenopus eggs. Proc Natl Acad Sci U S A. 1980 Jan;77(1):462–466. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kallenbach RJ. Continuous hypertonic conditions activate and promote the formation of new centrioles within cytasters in sea urchin eggs. Cell Biol Int Rep. 1982 Nov;6(11):1025–1031. [Abstract] [Google Scholar]
  • Kimelman D, Kirschner M, Scherson T. The events of the midblastula transition in Xenopus are regulated by changes in the cell cycle. Cell. 1987 Feb 13;48(3):399–407. [Abstract] [Google Scholar]
  • Kuriyama R, Borisy GG. Centriole cycle in Chinese hamster ovary cells as determined by whole-mount electron microscopy. J Cell Biol. 1981 Dec;91(3 Pt 1):814–821. [Europe PMC free article] [Abstract] [Google Scholar]
  • Labbe JC, Picard A, Peaucellier G, Cavadore JC, Nurse P, Doree M. Purification of MPF from starfish: identification as the H1 histone kinase p34cdc2 and a possible mechanism for its periodic activation. Cell. 1989 Apr 21;57(2):253–263. [Abstract] [Google Scholar]
  • Meijer L, Pondaven P. Cyclic activation of histone H1 kinase during sea urchin egg mitotic divisions. Exp Cell Res. 1988 Jan;174(1):116–129. [Abstract] [Google Scholar]
  • Miake-Lye R, Newport J, Kirschner M. Maturation-promoting factor induces nuclear envelope breakdown in cycloheximide-arrested embryos of Xenopus laevis. J Cell Biol. 1983 Jul;97(1):81–91. [Europe PMC free article] [Abstract] [Google Scholar]
  • Miki-Noumura T. Studies on the de novo formation of centrioles: aster formation in the activated eggs of sea urchin. J Cell Sci. 1977 Apr;24:203–216. [Abstract] [Google Scholar]
  • Minshull J, Blow JJ, Hunt T. Translation of cyclin mRNA is necessary for extracts of activated xenopus eggs to enter mitosis. Cell. 1989 Mar 24;56(6):947–956. [Abstract] [Google Scholar]
  • Mitchison T, Kirschner M. Microtubule assembly nucleated by isolated centrosomes. Nature. 1984 Nov 15;312(5991):232–237. [Abstract] [Google Scholar]
  • Murray AW, Kirschner MW. Cyclin synthesis drives the early embryonic cell cycle. Nature. 1989 May 25;339(6222):275–280. [Abstract] [Google Scholar]
  • Nagano H, Hirai S, Okano K, Ikegami S. Achromosomal cleavage of fertilized starfish eggs in the presence of aphidicolin. Dev Biol. 1981 Jul 30;85(2):409–415. [Abstract] [Google Scholar]
  • Newport J, Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. [Abstract] [Google Scholar]
  • Newport JW, Kirschner MW. Regulation of the cell cycle during early Xenopus development. Cell. 1984 Jul;37(3):731–742. [Abstract] [Google Scholar]
  • Phillips SG, Rattner JB. Dependence of centriole formation on protein synthesis. J Cell Biol. 1976 Jul;70(1):9–19. [Europe PMC free article] [Abstract] [Google Scholar]
  • Picard A, Harricane MC, Labbe JC, Doree M. Germinal vesicle components are not required for the cell-cycle oscillator of the early starfish embryo. Dev Biol. 1988 Jul;128(1):121–128. [Abstract] [Google Scholar]
  • Raff JW, Glover DM. Nuclear and cytoplasmic mitotic cycles continue in Drosophila embryos in which DNA synthesis is inhibited with aphidicolin. J Cell Biol. 1988 Dec;107(6 Pt 1):2009–2019. [Europe PMC free article] [Abstract] [Google Scholar]
  • Riabowol K, Draetta G, Brizuela L, Vandre D, Beach D. The cdc2 kinase is a nuclear protein that is essential for mitosis in mammalian cells. Cell. 1989 May 5;57(3):393–401. [Abstract] [Google Scholar]
  • Robbins E, Jentzsch G, Micali A. The centriole cycle in synchronized HeLa cells. J Cell Biol. 1968 Feb;36(2):329–339. [Europe PMC free article] [Abstract] [Google Scholar]
  • Sellitto C, Kuriyama R. Distribution of pericentriolar material in multipolar spindles induced by colcemid treatment in Chinese hamster ovary cells. J Cell Sci. 1988 Jan;89(Pt 1):57–65. [Abstract] [Google Scholar]
  • Sluder G, Lewis K. Relationship between nuclear DNA synthesis and centrosome reproduction in sea urchin eggs. J Exp Zool. 1987 Oct;244(1):89–100. [Abstract] [Google Scholar]
  • Sluder G, Miller FJ, Rieder CL. The reproduction of centrosomes: nuclear versus cytoplasmic controls. J Cell Biol. 1986 Nov;103(5):1873–1881. [Europe PMC free article] [Abstract] [Google Scholar]
  • Spiegelman BM, Lopata MA, Kirschner MW. Multiple sites for the initiation of microtubule assembly in mammalian cells. Cell. 1979 Feb;16(2):239–252. [Abstract] [Google Scholar]
  • Tuffanelli DL, McKeon F, Kleinsmith DM, Burnham TK, Kirschner M. Anticentromere and anticentriole antibodies in the scleroderma spectrum. Arch Dermatol. 1983 Jul;119(7):560–566. [Abstract] [Google Scholar]
  • Vandre DD, Davis FM, Rao PN, Borisy GG. Phosphoproteins are components of mitotic microtubule organizing centers. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4439–4443. [Europe PMC free article] [Abstract] [Google Scholar]
  • Vorobjev IA, Chentsov YuS Centrioles in the cell cycle. I. Epithelial cells. J Cell Biol. 1982 Jun;93(3):938–949. [Europe PMC free article] [Abstract] [Google Scholar]
  • Weisenberg RC, Rosenfeld AC. In vitro polymerization of microtubules into asters and spindles in homogenates of surf clam eggs. J Cell Biol. 1975 Jan;64(1):146–158. [Europe PMC free article] [Abstract] [Google Scholar]
  • Woodland HR, Adamson ED. The synthesis and storage of histones during the oogenesis of Xenopus laevis. Dev Biol. 1977 May;57(1):118–135. [Abstract] [Google Scholar]
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