Skip to main content
SpringerLink
Log in

Actin and myosin inhibitors block elongation of kinetochore fibre stubs in metaphase crane-fly spermatocytes

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary.

We used an ultraviolet microbeam to cut individual kinetochore spindle fibres in metaphase crane-fly spermatocytes. We then followed the growth of the “kinetochore stubs”, the remnants of kinetochore fibres that remain attached to kinetochores. Kinetochore stubs elongate with constant velocity by adding tubulin subunits at the kinetochore, and thus elongation is related to tubulin flux in the kinetochore microtubules. Stub elongation was blocked by cytochalasin D and latrunculin A, actin inhibitors, and by butanedione monoxime, a myosin inhibitor. We conclude that actin and myosin are involved in generating elongation and thus in producing tubulin flux in kinetochore microtubules. We suggest that actin and myosin act in concert with a spindle matrix to propel kinetochore fibres poleward, thereby causing stub elongation and generating anaphase chromosome movement in nonirradiated cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • LA Cameron G Yang D Cimini JC Canman O Kisurina-Evgenieva A Khodjakov G Danuser ED Salmon (2006) ArticleTitleKinesin 5-independent poleward flux of kinetochore microtubules in PtK1 cells J Cell Biol 173 173–179 Occurrence Handle16636143 Occurrence Handle10.1083/jcb.200601075 Occurrence Handle1:CAS:528:DC%2BD28XktVCgu74%3D

    Article  PubMed  CAS  Google Scholar 

  • W Chen D Zhang (2004) ArticleTitleKinetochore fibre dynamics outside the context of the spindle during anaphase Nat Cell Biol 6 227–231 Occurrence Handle15039774

    PubMed  Google Scholar 

  • BB Czaban A Forer AS Bajer (1993) ArticleTitleUltraviolet microbeam irradiation of chromosomal spindle fibres in Haemanthus katherinae endosperm. I. Behaviour of the irradiated region J Cell Sci 105 571–578 Occurrence Handle8408286

    PubMed  Google Scholar 

  • L Fabian A Forer (2005) ArticleTitleRedundant mechanisms for anaphase chromosome movements: crane-fly spermatocyte spindles normally use actin filaments but also can function without them Protoplasma 225 169–184 Occurrence Handle16228898 Occurrence Handle10.1007/s00709-005-0094-6

    Article  PubMed  Google Scholar 

  • A Forer (1965) ArticleTitleLocal reduction of spindle fiber birefringence in living Nephrotoma suturalis (Loew) spermatocytes induced by ultraviolet microbeam irradiation J Cell Biol 25 95–117 Occurrence Handle14342833 Occurrence Handle10.1083/jcb.25.1.95

    Article  PubMed  Google Scholar 

  • A Forer (1966) ArticleTitleCharacterization of the mitotic traction system, and evidence that birefringent spindle fibers neither produce nor transmit force for chromosome movement Chromosoma 19 44–98 Occurrence Handle5330243 Occurrence Handle10.1007/BF00332793 Occurrence Handle1:STN:280:CCiD3c3kslc%3D

    Article  PubMed  CAS  Google Scholar 

  • A Forer L Fabian (2005) ArticleTitleDoes 2,3-butanedione monoxime inhibit nonmuscle myosin? Protoplasma 225 1–4 Occurrence Handle15868207 Occurrence Handle10.1007/s00709-004-0077-z Occurrence Handle1:CAS:528:DC%2BD2MXjvVSqtb0%3D

    Article  PubMed  CAS  Google Scholar 

  • A Forer JD Pickett-Heaps (1998) ArticleTitleCytochalasin D and latrunculin affect chromosome behaviour during meiosis in crane-fly spermatocytes Chromosome Res 6 533–549 Occurrence Handle9886773 Occurrence Handle10.1023/A:1009224322399 Occurrence Handle1:CAS:528:DyaK1MXisVygsA%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • A Forer JD Pickett-Heaps (2005) ArticleTitleFibrin clots keep non-adhering living cells in place on glass for perfusion or fixation Cell Biol Int 29 721–730 Occurrence Handle16095930 Occurrence Handle10.1016/j.cellbi.2005.04.010 Occurrence Handle1:CAS:528:DC%2BD2MXpslymt7o%3D

    Article  PubMed  CAS  Google Scholar 

  • A Forer PJ Wilson (1994) ArticleTitleA model for chromosome movement during mitosis Protoplasma 179 95–105 Occurrence Handle10.1007/BF01403947

    Article  Google Scholar 

  • A Forer T Spurck JD Pickett-Heaps (1997) ArticleTitleUltraviolet microbeam irradiations of spindle fibres in crane-fly spermatocytes and newt epithelial cells: resolution of previously conflicting observations Protoplasma 197 230–240 Occurrence Handle10.1007/BF01288032

    Article  Google Scholar 

  • A Forer T Spurck JD Pickett-Heaps PJ Wilson (2003) ArticleTitleStructure of kinetochore fibres in crane-fly spermatocytes after irradiation with an ultraviolet microbeam: neither microtubules nor actin filaments remain in the irradiated region Cell Motil Cytoskeleton 56 173–192 Occurrence Handle14569597 Occurrence Handle10.1002/cm.10144

    Article  PubMed  Google Scholar 

  • NJ Ganem K Upton DA Compton (2005) ArticleTitleEfficient mitosis in human cells lacking poleward microtubule flux Curr Biol 15 1827–1832 Occurrence Handle16243029 Occurrence Handle10.1016/j.cub.2005.08.065 Occurrence Handle1:CAS:528:DC%2BD2MXhtFCjsL%2FM

    Article  PubMed  CAS  Google Scholar 

  • JR LaFountain SuffixJr CS Cohan AJ Siegel DJ LaFountain (2004) ArticleTitleDirect visualization of microtubule flux during metaphase and anaphase in crane-fly spermatocytes Mol Biol Cell 15 5724–5732 Occurrence Handle15469981 Occurrence Handle10.1091/mbc.E04-08-0750 Occurrence Handle1:CAS:528:DC%2BD2cXhtVGlsLfO

    Article  PubMed  CAS  Google Scholar 

  • H Maiato CL Rieder A Khodjakov (2004) ArticleTitleKinetochore-driven formation of kinetochore fibers contributes to spindle assembly during animal mitosis J Cell Biol 167 831–840 Occurrence Handle15569709 Occurrence Handle10.1083/jcb.200407090 Occurrence Handle1:CAS:528:DC%2BD2cXhtVOlsbrJ

    Article  PubMed  CAS  Google Scholar 

  • H Maiato A Khodjakov CL Rieder (2005) ArticleTitle Drosophila CLASP is required for the incorporation of microtubule subunits into fluxing kinetochore fibres Nat Cell Biol 7 42–47 Occurrence Handle15592460 Occurrence Handle10.1038/ncb1207 Occurrence Handle1:CAS:528:DC%2BD2MXlvFWi

    Article  PubMed  CAS  Google Scholar 

  • RL Margolis L Wilson (1978) ArticleTitleOpposite end assembly and disassembly of microtubules at steady state in vitro Cell 13 1–8 Occurrence Handle620419 Occurrence Handle10.1016/0092-8674(78)90132-0 Occurrence Handle1:CAS:528:DyaE1cXhtFKlsro%3D

    Article  PubMed  CAS  Google Scholar 

  • RL Margolis L Wilson (1981) ArticleTitleMicrotubule treadmills – possible molecular machinery Nature 203 705–711 Occurrence Handle10.1038/293705a0

    Article  Google Scholar 

  • TJ Mitchison (1989) ArticleTitlePolewards microtubule flux in the mitotic spindle: evidence from photoactivation of fluorescence J Cell Biol 109 637–652 Occurrence Handle2760109 Occurrence Handle10.1083/jcb.109.2.637 Occurrence Handle1:CAS:528:DyaL1MXltlSjtbk%3D

    Article  PubMed  CAS  Google Scholar 

  • DT Miyamoto ZE Perlman KS Burbank AC Groen TJ Mitchison (2004) ArticleTitleThe kinesin Eg5 drives poleward microtubule flux in Xenopus laevis egg extract spindles J Cell Biol 167 813–818 Occurrence Handle15583027 Occurrence Handle10.1083/jcb.200407126 Occurrence Handle1:CAS:528:DC%2BD2cXhtVOlsbrP

    Article  PubMed  CAS  Google Scholar 

  • JD Pickett-Heaps A Forer T Spurck (1996) ArticleTitleRethinking anaphase: where “Pac-Man” fails and why a role for the spindle matrix is likely Protoplasma 192 1–10 Occurrence Handle10.1007/BF01273239

    Article  Google Scholar 

  • JD Pickett-Heaps A Forer T Spurck (1997) ArticleTitleTraction fibre: toward a “tensegral” model of the spindle Cell Motil Cytoskeleton 37 1–6 Occurrence Handle9142434 Occurrence Handle10.1002/(SICI)1097-0169(1997)37:1<1::AID-CM1>3.0.CO;2-D Occurrence Handle1:STN:280:ByiB1cjjsVY%3D

    Article  PubMed  CAS  Google Scholar 

  • GC Rogers SL Rogers TA Schwimmer SC Ems-McClung CE Walczak RD Vale JM Scholey DJ Sharp (2004) ArticleTitleTwo mitotic kinesins cooperate to drive sister chromatid separation during anaphase Nature 427 364–370 Occurrence Handle14681690 Occurrence Handle10.1038/nature02256 Occurrence Handle1:CAS:528:DC%2BD2cXltFCgtQ%3D%3D

    Article  PubMed  CAS  Google Scholar 

  • GC Rogers SL Rogers DJ Sharp (2005) ArticleTitleSpindle microtubules in flux J Cell Sci 118 1105–1116 Occurrence Handle15764594 Occurrence Handle10.1242/jcs.02284 Occurrence Handle1:CAS:528:DC%2BD2MXjs1Chs7k%3D

    Article  PubMed  CAS  Google Scholar 

  • JM Scholey GC Rogers DJ Sharp (2001) ArticleTitleMitosis, microtubules, and the matrix J Cell Biol 154 261–266 Occurrence Handle11470815 Occurrence Handle10.1083/jcb.200101097 Occurrence Handle1:CAS:528:DC%2BD3MXlsVOjs7g%3D

    Article  PubMed  CAS  Google Scholar 

  • PJ Sillers A Forer (1983) ArticleTitleAction spectrum for changes in spindle fibre birefringence after ultraviolet microbeam irradiations of single chromosomal spindle fibres in crane-fly spermatocytes J Cell Sci 62 1–25 Occurrence Handle6619201 Occurrence Handle1:STN:280:BiuD3cfjt1w%3D

    PubMed  CAS  Google Scholar 

  • RV Silverman-Gavrila A Forer (2000) ArticleTitleEvidence that actin and myosin are involved in the poleward flux of tubulin in metaphase kinetochore microtubules of crane-fly spermatocytes J Cell Sci 113 597–609 Occurrence Handle10652253 Occurrence Handle1:CAS:528:DC%2BD3cXhvFSgtr8%3D

    PubMed  CAS  Google Scholar 

  • T Spurck A Forer JD Pickett-Heaps (1997) ArticleTitleUltraviolet microbeam irradiations of epithelial and spermatocyte spindles suggest that forces act on the kinetochore fibre and are not generated by its disassembly Cell Motil Cytoskeleton 36 136–148 Occurrence Handle9015202 Occurrence Handle10.1002/(SICI)1097-0169(1997)36:2<136::AID-CM4>3.0.CO;2-7 Occurrence Handle1:STN:280:ByiC28jlvVA%3D

    Article  PubMed  CAS  Google Scholar 

  • PJ Wilson A Forer (1989a) ArticleTitleThe behaviour of microtubules in chromosomal spindle fibres irradiated singly or doubly with ultraviolet light J Cell Sci 94 625–634

    Google Scholar 

  • PJ Wilson A Forer (1989b) ArticleTitleIdentifying the site of microtubule polymerization during regrowth of UV-sheared kinetochore fibres using antibodies against acetylated alpha-tubulin Cell Biol Int Rep 13 823–832 Occurrence Handle10.1016/0309-1651(89)90123-9 Occurrence Handle1:STN:280:By%2BD3s%2FksVI%3D

    Article  CAS  Google Scholar 

  • PJ Wilson A Forer (1997) ArticleTitleEffects of nanomolar taxol on crane-fly spermatocyte spindles indicate that acetylation of kinetochore microtubules can be used as a marker of poleward tubulin flux Cell Motil Cytoskeleton 37 20–32 Occurrence Handle9142436 Occurrence Handle10.1002/(SICI)1097-0169(1997)37:1<20::AID-CM3>3.0.CO;2-L Occurrence Handle1:CAS:528:DyaK2sXjtleqtLw%3D

    Article  PubMed  CAS  Google Scholar 

  • PJ Wilson A Forer C Leggiadro (1994) ArticleTitleEvidence that kinetochore microtubules in crane-fly spermatocytes disassemble during anaphase primarily at the poleward end J Cell Sci 107 3015–3027 Occurrence Handle7699001 Occurrence Handle1:CAS:528:DyaK2MXisFSksbc%3D

    PubMed  CAS  Google Scholar 

  • D Zhang RB Nicklas (1996) ArticleTitle‘Anaphase’ and cytokinesis in the absence of chromosomes Nature 382 466–468 Occurrence Handle8684488 Occurrence Handle10.1038/382466a0 Occurrence Handle1:CAS:528:DyaK28Xks1Olurc%3D

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biology Department, York University, Toronto, Ontario

    A. Forer

  2. School of Botany, Melbourne University, Parkville, Victoria

    T. Spurck & J. D. Pickett-Heaps

Authors
  1. A. Forer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Spurck
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. D. Pickett-Heaps
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. Forer.

Additional information

Correspondence: A. Forer, Biology Department, York University, 4700 Keele Street, Toronto, ON M3J 1P3, Canada.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Forer, A., Spurck, T. & Pickett-Heaps, J. Actin and myosin inhibitors block elongation of kinetochore fibre stubs in metaphase crane-fly spermatocytes. Protoplasma 232, 79–85 (2007). https://doi.org/10.1007/s00709-007-0265-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00709-007-0265-8

Search

Navigation