Europe PMC
Nothing Special   »   [go: up one dir, main page]

Europe PMC requires Javascript to function effectively.

Either your web browser doesn't support Javascript or it is currently turned off. In the latter case, please turn on Javascript support in your web browser and reload this page.

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Polarity of microtubules nucleated by centrosomes and chromosomes of Chinese hamster ovary cells in vitro.

The Journal of Cell Biology, 01 Jan 1980, 84(1):151-159
https://doi.org/10.1083/jcb.84.1.151 PMID: 7350167 PMCID: PMC2110533

This article is in the Europe PMC Open access subset. Refer to the copyright information in the article for licensing details.
Free full text in Europe PMC

Abstract 

The structural and growth polarities of centrosomal and chromosomal microtubules were studied by analyzing the kinetics of growth of these microtubules and those initiated by flagellar seeds. By comparing rates of elongation of centrosomal and flagellar-seeded microtubules, we determined whether the centrosomal microtubules were free to grow at their plus ends only, minus ends ony, or at both ends. Our results show that centrosomal microtubules elongate at a rate corresponding to the addition of subunits at the plus end only. The depolymerization rate was also equivalent to that for the plus end only. Chromosomal microtubule elongation was similar to the centrosome-initiated growth. Since the data do not support the hypothesis that both ends of these spindle microtubules are able to interact with monomer in solution, then growth must occur only distal or only proximal to the organizing centers, implying tha the opposite ends in unavailable for exchange of subunits. Experiments with flagellar-seeded microtubules serving as internal controls indicated that the inactivity of the minus end could not be accounted for by a diffusible inhibitor, suggesting a structural explanation. Since there is no apparent way in which the distal ends may be capped, whereas the proximal ends are embedded in the pericentriolar cloud, we conclude that centrosomal microtubules are oriented with their plus ends distal to the site of nucleation. A similar analysis for chromosomal microtubules suggests that they too must be oriented with their plus ends distal to the site of initiation.

Free full text 

Logo of jcellbiolLink to Publisher's site
J Cell Biol. 1980 Jan 1; 84(1): 151–159.
PMCID: PMC2110533
PMID: 7350167

Polarity of microtubules nucleated by centrosomes and chromosomes of Chinese hamster ovary cells in vitro

LG Bergen, R Kuriyama, and GG Borisy
Copyright and License information Disclaimer

Abstract

The structural and growth polarities of centrosomal and chromosomal microtubules were studied by analyzing the kinetics of growth of these microtubules and those initiated by flagellar seeds. By comparing rates of elongation of centrosomal and flagellar-seeded microtubules, we determined whether the centrosomal microtubules were free to grow at their plus ends only, minus ends ony, or at both ends. Our results show that centrosomal microtubules elongate at a rate corresponding to the addition of subunits at the plus end only. The depolymerization rate was also equivalent to that for the plus end only. Chromosomal microtubule elongation was similar to the centrosome-initiated growth. Since the data do not support the hypothesis that both ends of these spindle microtubules are able to interact with monomer in solution, then growth must occur only distal or only proximal to the organizing centers, implying tha the opposite ends in unavailable for exchange of subunits. Experiments with flagellar-seeded microtubules serving as internal controls indicated that the inactivity of the minus end could not be accounted for by a diffusible inhibitor, suggesting a structural explanation. Since there is no apparent way in which the distal ends may be capped, whereas the proximal ends are embedded in the pericentriolar cloud, we conclude that centrosomal microtubules are oriented with their plus ends distal to the site of nucleation. A similar analysis for chromosomal microtubules suggests that they too must be oriented with their plus ends distal to the site of initiation.

Full Text

The Full Text of this article is available as a PDF (823K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Summers K, Kirschner MW. Characteristics of the polar assembly and disassembly of microtubules observed in vitro by darkfield light microscopy. J Cell Biol. 1979 Oct;83(1):205–217. [Europe PMC free article] [Abstract] [Google Scholar]
  • Allen C, Borisy GG. Structural polarity and directional growth of microtubules of Chlamydomonas flagella. J Mol Biol. 1974 Dec 5;90(2):381–402. [Abstract] [Google Scholar]
  • Bajer AS. Interaction of microtubules and the mechanism of chromosome movement (zipper hypothesis). 1. General principle. Cytobios. 1973 Nov;8(31):139–160. [Abstract] [Google Scholar]
  • Bergen LG, Borisy GG. Head-to-tail polymerization of microtubules in vitro. Electron microscope analysis of seeded assembly. J Cell Biol. 1980 Jan;84(1):141–150. [Europe PMC free article] [Abstract] [Google Scholar]
  • Borisy GG, Marcum JM, Olmsted JB, Murphy DB, Johnson KA. Purification of tubulin and associated high molecular weight proteins from porcine brain and characterization of microtubule assembly in vitro. Ann N Y Acad Sci. 1975 Jun 30;253:107–132. [Abstract] [Google Scholar]
  • Borisy GG. Polarity of microtubules of the mitotic spindle. J Mol Biol. 1978 Sep 25;124(3):565–570. [Abstract] [Google Scholar]
  • Dietz R. Die Assembly-Hypothese der Chromosomenbewegung und die Veränderungen der Spindellänge während der Anaphase I in Spermatocyten von Pales ferruginea (Tipulidae, Diptera. Chromosoma. 1972;38(1):11–76. [Abstract] [Google Scholar]
  • FORER A. LOCAL REDUCTION OF SPINDLE FIBER BIREFRINGENCE IN LIVING NEPHROTOMA SUTURALIS (LOEW) SPERMATOCYTES INDUCED BY ULTRAVIOLET MICROBEAM IRRADIATION. J Cell Biol. 1965 Apr;25:SUPPL–SUPPL117. [Europe PMC free article] [Abstract] [Google Scholar]
  • Gould RR, Borisy GG. The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation. J Cell Biol. 1977 Jun;73(3):601–615. [Europe PMC free article] [Abstract] [Google Scholar]
  • Gould RR, Borisy GG. Quantitative initiation of microtubule assembly by chromosomes from Chinese hamster ovary cells. Exp Cell Res. 1978 May;113(2):369–374. [Abstract] [Google Scholar]
  • Hyams JS, Borisy GG. Nucleation of microtubules in vitro by isolated spindle pole bodies of the yeast Saccharomyces cerevisiae. J Cell Biol. 1978 Aug;78(2):401–414. [Europe PMC free article] [Abstract] [Google Scholar]
  • Inoué S, Ritter H., Jr Dynamics of mitotic spindle organization and function. Soc Gen Physiol Ser. 1975;30:3–30. [Abstract] [Google Scholar]
  • Inoué S, Ritter H., Jr Mitosis in Barbulanympha. II. Dynamics of a two-stage anaphase, nuclear morphogenesis, and cytokinesis. J Cell Biol. 1978 Jun;77(3):655–684. [Europe PMC free article] [Abstract] [Google Scholar]
  • Margolis RL, Wilson L, Keifer BI. Mitotic mechanism based on intrinsic microtubule behaviour. Nature. 1978 Mar 30;272(5652):450–452. [Abstract] [Google Scholar]
  • McGill M, Brinkley BR. Human chromosomes and centrioles as nucleating sites for the in vitro assembly of microtubules from bovine brain tubulin. J Cell Biol. 1975 Oct;67(1):189–199. [Europe PMC free article] [Abstract] [Google Scholar]
  • McIntosh JR, Cande WZ, Snyder JA. Structure and physiology of the mammalian mitotic spindle. Soc Gen Physiol Ser. 1975;30:31–76. [Abstract] [Google Scholar]
  • Mc2ntosh JR, Cande Z, Snyder J, Vanderslice K. Studies on the mechanism of mitosis. Ann N Y Acad Sci. 1975 Jun 30;253:407–427. [Abstract] [Google Scholar]
  • Nicklas RB. Mitosis. Adv Cell Biol. 1971;2:225–297. [Abstract] [Google Scholar]
  • Nicklas RB, Brinkley BR, Pepper DA, Kubai DF, Rickards GK. Electron microscopy of spermatocytes previously studied in life: methods and some observations on micromanipulated chromosomes. J Cell Sci. 1979 Feb;35:87–104. [Abstract] [Google Scholar]
  • Oakley BR, Heath IB. The arrangement of microtubules in serially sectioned spindles of the alga Cryptomonas. J Cell Sci. 1978 Jun;31:53–70. [Abstract] [Google Scholar]
  • Pickett-Heaps JD, Tippit DH. The diatom spindle in perspective. Cell. 1978 Jul;14(3):455–467. [Abstract] [Google Scholar]
  • Snyder JA, McIntosh JR. Initiation and growth of microtubules from mitotic centers in lysed mammalian cells. J Cell Biol. 1975 Dec;67(3):744–760. [Europe PMC free article] [Abstract] [Google Scholar]
  • Subirana JA. Role of spindle microtubules in mitosis. J Theor Biol. 1968 Jul;20(1):117–123. [Abstract] [Google Scholar]
  • Telzer BR, Moses MJ, Rosenbaum JL. Assembly of microtubules onto kinetochores of isolated mitotic chromosomes of HeLa cells. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4023–4027. [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]
Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

Full text links 

Read article at publisher's site: https://doi.org/10.1083/jcb.84.1.151

Read article for free, from open access legal sources, via Unpaywall: http://jcb.rupress.org/content/84/1/151.full.pdfUnpaywall

Citations & impact 

Impact metrics

80
Citations
Jump to Citations

Citations of article over time

Article citations

Go to all (80) article citations

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.