Hostname: page-component-77c89778f8-9q27g Total loading time: 0 Render date: 2024-07-22T15:08:43.388Z Has data issue: false hasContentIssue false
  • English
  • Français

Activation-dependent and activation-independent localisation of calmodulin to the mitotic apparatus during the first cell cycle of the Lytechinus piçtus embryo.

Published online by Cambridge University Press:  26 September 2008

Martin Wilding ,
Katalin Török  and
Michael Whitaker
Martin Wilding*
Affiliation:
Department of Physiology, University College London, London, UK.
Katalin Török
Affiliation:
Department of Physiology, University College London, London, UK.
Michael Whitaker
Affiliation:
Department of Physiology, University College London, London, UK.
*
Michael Whitaker, Department of Physiological Sciences, University of Newcastle, Medical School, Newcastle upon Tyne NE2 4HH, UK.
Get access Rights & Permissions [Opens in a new window]

Summary

We have used confocal microscopy and a fluorescent calmodulin probe to examine the mechanism of localisation of calmodulin during the first cell cycle of the sea urchin zygote. Using fluoresceincalmodulin, calmodulin can be observed within the nucleus and interphase astral microtubule arrays as cells approach mitosis. During mitosis, calmodulin redistributes to the mitotic apparatus and to condensed chromosomes. Quantitative analysis with reference to a control dye (fluorescein-dextran) shows that the distribution of calmodulin is specific. We used a competitive inhibitor of calcium-dependent calmodulin binding (Trp-peptide; Török & Trentham (1994) Biochemistry 33, 12807–20) to test whether the cell cycle localisation of calmodulin was due to its binding to targets on activation. The Trp-peptide eliminates localisation of calmodulin within the nucleus. However, microtubule localisation persists in the presence of the Trp-peptide. These data show that calmodulin can localise by calcium (and hence activation)-dependent as well as calcium-independent mechanisms. This suggests that distinct mechanisms of localisation may be involved in the regulation of the differential functions of calmodulin, at least during the cell cycle.

Keywords

CalciumCalmodulinFluorescenceMitosisTa-calmodulim
Type
Article
Information
Zygote , Volume 3 , Issue 3 , August 1995 , pp. 219 - 224
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Andersen, B., Osborn, M., Weber, M. (1978). Specific visualisation of the distribution of the calcium-dependent regulatory protein of cyclic nucleotide phosphodiesterase (modulator protein) in tissue culture cells by immunofluorescence microscopy: mitosis and intracellular bridge. Cytobiologie 17, 354–64.Google Scholar
Anraku, Y., Ohya, Y., & Iida, H. (1991). Cell cycle control by calcium and calmodulin in Saccharomyces cerevisiae. Biochim. Biophys. Acta. 1093, 169–77.Google Scholar
Ciapa, B., Pesando, D., Wilding, M. & Whitaker, M.J. (1994). Cell cycle calcium transients driven by cyclic changes in inositol triphosphate levels. Nature 368, 875–8.Google Scholar
Cohen, J., & Klee, C. (1988). Calmodulin. Amsterdam: Elsevier.Google Scholar
Colca, J.R., Dewald, D.B., Pearson, J.D., Palazuk, B.J., Laurino, J.P. & MacDonald, J.M. (1987). Insulin stimulates the phosphorylation of calmodulin in intact adipocytes. J. Biol. chem. 262, 11399–402.Google Scholar
Gough, A., & Taylor, L. (1993). Fluorescence anisotropy imaging maps calmodulin binding during cellular contraction and locomotion. J. Cell Biol. 121, 1095–107.Google Scholar
Graff, J.M., Young, T.N., Johnson, J.D. & Blackshear, P.J. (1989). Phosphorylation-regulated calmodulin binding to a prominent cellular substrate for protein kinase C. J. Biol. Chem. 264 21818–23.Google Scholar
Hamaguchi, Y., & Iwasa, F. (1980). Localisation of fluorescently labelled calmodulin in living sea urchin eggs during early development. Biomed. Res. 1, 502–9.Google Scholar
Klee, C., Crouch, T., & Richman, P. (1980). Calmodulin. Annu. Rev. Biochem. 49, 489–515.Google Scholar
Lorca, T., Galas, S., Fesquet, D., Devault, A., Cavadore, J., & Dorée, M. (1991). Degradation of the proto-oncogene product p39mos is not necessary for cyclin proteolysis and exit from meiotic metaphase: requirement for a calcium/calmodulin-dependent step. EMBO J. 10, 2087–93.Google Scholar
Lorca, T., Cruzalegui, F., Fesquet, D., Cavadore, J., Méry, J., Means, A., & Dorée, M. (1993). Calmodulin-dependent protein kinase II mediates inactivation of MPF and CSF upon fertilisation of Xenopus eggs. Nature 366, 270–3.CrossRefGoogle ScholarPubMed
Lu, K., & Means, A. (1993). Regulation of the cell cycle by calcium and calmodulin. Endocr. Rev. 14, 40–58.Google Scholar
Means, A. (1988). Molecular mechanisms of action of calmodulin. Rec. Prog. Hormone Res. 44, 223–62.Google Scholar
Means, A. (1994). Calcium, calmodulin and cell cycle regulation. FEBS Lett. 347, 1–4.CrossRefGoogle ScholarPubMed
Means, A., & Dedman, J. (1980). Calmodulin: an intracellular calcium receptor. Nature 285, 73–7.CrossRefGoogle ScholarPubMed
Morin, N.Abrieu, A., Lorca, T., Martin, F., & Dorée, M. (1994). The proteolysis-dependent metaphase to anaphase transition: calcium/calmodulin-dependent protein kinase II mediates onset of anaphase in extracts prepared from unfertilised Xenopus eggs. EMBO J. 13, 4343–52.Google Scholar
Ohya, Y., & Anraku, Y. (1992). Yeast calmodulin: structural and functional elements essential for the cell cycle. Cell Calcium 13, 445–55.Google Scholar
Ohya, Y., & Botstein, D. (1994). Diverse essential functions revealed by complementing yeast calmodulin mutants. Science 263, 963–6.Google Scholar
Portolés, M., Faura, M., Renau-Piqueras, J., Iborra, F., Saez, J., Guerri, C., Serratosa, J., Rius, E., & Bachs, O. (1994). Nuclear calmodulin/62 kDa calmodulin binding protein complexes in interphase and mitotic cells. J. Cell Sci. 107, 3601–14.Google Scholar
Rasmussen, C., & Means, A. (1989). Calmodulin, cell growth and gene expression. Trends Neurosci. 11, 433–8.Google Scholar
Saville, M.K., & Houslay, M.D. (1994). Phosphorylation of calmodulin on Tyr99 selectively attenuates the action of calmodulin antagonists on type 1 cyclic nucleotide phosphodiesterase. Biochem J. 299, 863–5.Google Scholar
Sweet, S., Rogers, C., & Welsh, M. (1988). Calmodulin stabilisation of kinetochore microtubule structure to the effect of nocodazole. J. Cell Biol. 107, 2243–51.CrossRefGoogle Scholar
Terasaki, M., & Jaffe, L. (1991). Organisation of the sea urchin endoplasmic reticulum and its reorganisation at fertilisation. J. Cell Biol. 114, 929–40.Google Scholar
Török, K. & Trentham, D. (1994). Mechanism of 2-chloro-(epsilon-amino-Lys75)-[6-[4-(n,n'-diethylamino)phenyl]-1,3,5-triazin-4-yl]calmodulin interactions with smooth muscle myosin light chain kinase and derived peptides. Biochemistry 33, 12807–20.CrossRefGoogle Scholar
Török, K., & Whitaker, M. (1994). Taking a long, hard look at calmodulin's warm embrace. BioEssays 16, 221–4.Google Scholar
Török, K., Cowley, D., Brandmeier, B., Howell, S., Aitken, A. & Trentham, D. (1995). Inhibition of steady state activity of smooth muscle myosin light chain kinase by calmodulin binding peptides and fluorescent calmodulin derivatives. Unpublished observations.Google Scholar
Vantard, M., Lambert, A., de, Mey J, Picquot, P. & Van, Eldick L. (1985). Characterisation and immunocytochemical distribution of calmodulin in higher plant endosperm cells: localisation in the mitotic apparatus. J. Cell Biol. 101, 488–99.Google Scholar
Welsh, M., Dedman, J., Brinkley, B., & Means, A. (1978). Calcium-dependent regulatory protein: localisation in the mitotic apparatus of eukaryotic cells. Proc. Natl. Acad. Sci., USA 75, 1867–71.Google Scholar
Welsh, M., Dedman, J., Brinkley, B., & Means, A. (1979). Tubulin and calmodulin: effects of microtubule and microfilament inhibitors on localisation in the mitotic apparatus. J. Cell Biol. 81, 624–34.Google Scholar
Whitaker, M. (1995). Regulation of the cell division cycle by inositol trisphosphate and the calcium signalling pathway. Adv. Sec. Mess. Phospho. Res. 30, 299–310.CrossRefGoogle Scholar
Whitaker, M.J., & Patel, R. (1990). Calcium and cell cycle control. Development 108, 525–42.Google Scholar
Wilding, M., Wright, E., Patel, R., & Whitaker, M. (1995). Spatial characteristics of calcium transients associated with mitosis entry in early sea urchin embryos. Unpublished observations.Google Scholar
Zavortink, M., Welsh, M., & McIntosh, J. (1983). The distribution of calmodulin in living mitotic cells. Exp. Cell Res. 149, 375–85.Google Scholar