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Cell cycle dependent distribution of a centrosomal antigen at the perinuclear MTOC or at the kinetochores of higher plant cells

Abstract

Compelling evidence has been obtained in favour of the idea that the nuclear surface of higher plant cells is a microtubule-nucleating and/or organizing site (MTOC), in the absence of defined centrosomes. How these plant MTOC proteins are redistributed and function during the progression of the cell cycle remains entirely unknown. Using a monoclonal antibody (mAb 6C6) raised against isolated calf thymus centrosomes and showing apparent reaction with the plant nuclear surface, we followed the targeted antigen distribution during mitosis and meiosis of higher plants. Immunoblot analysis of protein fractions from Allium root meristematic cell extracts probed with mAb 6C6 reveals a polypeptide of an apparent Mr of 78000. In calf centrosome extracts, a polypeptide of comparable molecular mass is found in addition to a major antigen of Mr 180000 after mAb 6C6 immunoblotting. During mitotic initiation, the plant antigen is prominent on the periphery of the prophase nucleus. When the nuclear envelope breaks down, the antigen suddenly becomes associated with the centromere-kinetochores until late anaphase. In telophase, when the nuclear envelope is being reconstructed, it is no longer detected at the kinetochores but is solely associated again with the nuclear surface. This antigen displays a unique spatial and temporal distribution, which may reflect the pathway of plant protein(s) between the nuclear surface and the kinetochores under cell cycle control. So far, such processes have not been described in higher plant cells. These observations shed light on the putative activity of the plant kinetochore as a protein transporter. They also suggest that a plant centrosome-like antigen may have different cytoskeletal related functions depending on cell cycle regulated changes in its subcellular distribution.

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Abbreviations

mAb:

monoclonal antibody

MSB:

microtubule stabilizing buffer

TBS:

Tris buffered saline

MTOC:

microtubule organizing centre

References

  1. Bajer AS, Mole-Bajer J (1986) Reorganization of microtubules in endosperm cells and cell fragments of the higher plant Haemanthus in vivo. J Cell Biol 102:263–281

  2. Bloom K (1993) The centromere frontier: kinetochore components, microtubule-based motility, and the CEN-Value-paradox. Cell 73:621–624

  3. Baskin TI, Cande WZ (1990) The structure and function of the mitotic spindle in flowering plants. Annu Rev Plant Physiol Plant Mol Biol 41:277–315

  4. Brinkley BR, Ouspenski I, Zinkowski RP (1992) Structure and molecular organization of the centromere-kinetochore complex. Trends Cell Biol 2:15–21

  5. Calarco-Guillan PP, Siebert MC, Hubbie R, Mitchison T, Kirschner M (1983) Centrosome development in early mouse embryos as defined by an autoantibody against pericentriolar material. Cell 35:621–629

  6. Chevrier V, Komesli S, Schmitt AC, Vantard M, Lambert AM, Job D (1992) A monoclonal antibody, raised against mammalian centrosomes and screened by recognition of plant microtubule organizing centers, identifies a pericentriolar component in different cell types. J Cell Sci 101:823–835

  7. Clayton LC, Black CM, Lloyd CW (1985) Microtubule nucleating sites in higher plant cells identified by an autoantibody against pericentriolar material. J Cell Biol 101:319–324

  8. Compton DA, Yen TJ, Cleveland DW (1991) Identification of novel centromere/kinetochore associated proteins using monoclonal antibodies generated against human mitotic chromosome scaffolds. J Cell Biol 112:1083–1097

  9. Earnshaw WC, Bernat RJ (1991) Chromosomal passengers: towards an integrated view of mitosis. Chromosoma 100:139–146

  10. Earnshaw WC, Cooke CA (1991) Analysis of the distribution of the INCENPs throughout mitosis reveals the existence of a pathway of structural changes in the chromosome during metaphase and early events in cleavage furrow formation. J Cell Sci 98:443–461

  11. Harper JDI, Mitchison JM; Williamson RE, John PCL (1989) Does the autoimmune serum 5051 specifically recognize microtubule organizing centres in higher plant cells? Cell Biol Int Rep 13:471–483

  12. Joshi HC, Palacios MJ, McNamura L, Cleveland DW (1992) γ-tubulin is a centrosomal protein required for cell cycle dependent microtubule nucleation. Nature 356:80–831

  13. Kellog DR, Field CM, Alberts BM (1989) Identification of microtubule-associated protein in the centrosome, spindle and kinetochore of the early drosophila embryo. J Cell Biol 109:2977–2991

  14. Lambert AM (1993) Microtubule-organizing centers in higher plants. Curr Opin Cell Biol 5:116–122

  15. Lambert AM, Lloyd CW (1994) The higher plant microtubule cycle. In: Hyams JS, Lloyd CW (eds) Microtubules. Wiley Liss, New York, pp 325–341

  16. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

  17. Liu B, Palevitz BA (1991) Kinetochore fiber formation in dividing generative cells of Tradescantia. J Cell Sci 98:475–482

  18. Liu B, Marc J, Joshi HC, Palevitz BA (1993) A γ-tubulin-related protein associated with the microtubule arrays of higher plants in a cell cycle-dependent manner. J Cell Sci 104:1217–1228

  19. McDonald AR, Liu B, Joshi HC, Palevitz BA (1993) γ-tubulin is associated with a cortical-microtubule-organizing zone in the developing guard cells of Allium cepa. Planta 191:357–361

  20. Melan M, Sluder G (1992) Redistribution and differential extraction of soluble proteins in permeabilized cultured cells: implications for immunofluorescence microscopy. J Cell Sci 101:731–743

  21. Mole-Bajer J, Bajer AS, Zinkowski RP, Balczon RD, Brinkley BR (1990) Autoantibodies from a patient with scleroderma CREST recognized kinetochores of the higher plant Heamanthus. Proc Natl Acad Sci USA 87:3599–3603

  22. Morejohn LC (1991) The molecular pharmacology of plant tubulin and microtubules. In: Lloyd CW (ed) The cytoskeletal basis of plant growth and form. Academic Press, London New York, pp 29–43

  23. Moroi Y, Peebles C, Fritzler MJ, Steigerwald J, Tan EM (1980) Autoantibodies to centromere (kinetochore) in scleroderma sera. Proc Natl Acad Sci USA 77:1627–1631

  24. Oakley BR (1992) γ-tubulin: the microtubule organizer. Trends Cell Biol 2:1–5

  25. Palevitz BA (1990) Kinetochore behavior during generative cell division in Tradescantia virginiana. Protoplasma 157:120–127

  26. Palevitz BA (1993) Morphological plasticity of the mitotic apparatus in plants and its developmental consequences. Plant Cell 5:1001–1009 (Review)

  27. Rattner JB, Wang T, Mack G, Fritzler MJ, Martin L, Valencia D (1992) MSA-36: A chromosomal and mitotic spindle-associated protein. Chromosoma 101:625–633

  28. Rieder CL (1990) Formation of the astral mitotic spindle: ultrastructural basis for the centrosome-kineotochore interaction. Electron Microsc Rev 3:269–300

  29. Schmit AC, Vantard M, DeMey J, Lambert AM (1983) Aster-like microtubule centers establish spindle polarity during interphase-mitosis transition. Plant Cell Rep 2:285–288

  30. Smirnova EA; Bajer AS (1992) Spindle poles in higher plant mitosis. Cell Motil Cytoskeleton 23:1–7

  31. Stoppin V, Vantard M, Schmitt AC, Lambert AM (1994) Isolated maize nuclei can nucleate microtubule assembly: the nuclear surface in higher plants has a centrosome-like activity. Plant Cell 6:1099–1106

  32. Tousson A, Zeng C, Brinkley BR, Valdivia MM (1991) Centrophilin: a novel mitotic spindle protein involved in microtubule nucleation. J Cell Biol 112:427–440

  33. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

  34. Vantard M, Levilliers N, Hill AM, Adoutte A, Lambert AM (1990) Incorporation of Paramecium axonemal tubulin into higher plant cells reveals functional sites of microtubule assembly. Proc Natl Acad Sci USA 87:8825–8829

  35. Wick SM (1985) The higher plant mitotic apparatus: redistribution of microtubules, calmodulin and microtubule initiation material during its establishment. Cytobios 43:285–294

  36. Yen TJ, Compton TA, Wise D, Zinkowski RP, Brinkley BR, Earnshaw WC, Cleveland DW (1991) CENP E, a novel human centromere-associated protein required for progression from metaphase to anaphase. EMBO J 10:1245–1254

  37. Zhang DH, Wadsworth P, Hepler PK (1990) Microtubule dynamics in living cells: confocal imaging of microinjected fluorescent brain tubulin. Proc Natl Acad Sci USA 87:8820–8824

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Schmit, A., Stoppin, V., Chevrier, V. et al. Cell cycle dependent distribution of a centrosomal antigen at the perinuclear MTOC or at the kinetochores of higher plant cells. Chromosoma 103, 343–351 (1994). https://doi.org/10.1007/BF00417882

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Keywords

  • Cell Cycle
  • Nuclear Envelope
  • Cell Cycle Control
  • Subcellular Distribution
  • Meristematic Cell