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Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap.

Publication ,  Journal Article
Voter, WA; O'Brien, ET; Erickson, HP
Published in: Cell Motil Cytoskeleton
1991

Microtubules were assembled from purified tubulin in the buffer originally used to study dynamic instability (100 mM PIPES, 2 mM EGTA, 1 mM magnesium, 0.2 mM GTP) and then diluted in the same buffer to study the rate of disassembly. Following a 15-fold dilution, microtubule polymer decreased linearly to about 20% of the starting value in 15 sec. We determined the length distribution of microtubules before dilution, and prepared computer simulations of polymer loss for different assumed rates of disassembly. Our experimental data were consistent with a disassembly rate per microtubule of 60 microns/min. This is the total rate of depolymerization for microtubules in the rapid shortening phase, as determined by light microscopy of individual microtubules (Walker et al.: Journal of Cell Biology 107:1437-1448, 1988). We conclude, therefore, that microtubules began rapid shortening at both ends upon dilution. Moreover, since we could detect no lag between dilution and the onset of rapid disassembly, the transition from elongation to rapid shortening apparently occurred within 1 sec following dilution. Assuming that this transition (catastrophe) involves the loss of the GTP cap, and that cap loss is achieved by the sequential dissociation of GTP-tubulin subunits following dilution, we can estimate the maximum size of the cap based on the kinetic data and model interpretation of Walker et al. The cap is probably shorter than 40 and 20 subunits at the plus and minus ends, respectively.

Duke Scholars

Published In

Cell Motil Cytoskeleton

DOI

ISSN

0886-1544

Publication Date

1991

Volume

18

Issue

1

Start / End Page

55 / 62

Location

United States

Related Subject Headings

  • Tubulin
  • Solutions
  • Microtubules
  • Kinetics
  • Guanosine Triphosphate
  • Developmental Biology
  • Animals
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences
  • 0601 Biochemistry and Cell Biology
 

Citation

APA
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ICMJE
MLA
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Voter, W. A., O’Brien, E. T., & Erickson, H. P. (1991). Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap. Cell Motil Cytoskeleton, 18(1), 55–62. https://doi.org/10.1002/cm.970180106
Voter, W. A., E. T. O’Brien, and H. P. Erickson. “Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap.Cell Motil Cytoskeleton 18, no. 1 (1991): 55–62. https://doi.org/10.1002/cm.970180106.
Voter WA, O’Brien ET, Erickson HP. Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap. Cell Motil Cytoskeleton. 1991;18(1):55–62.
Voter, W. A., et al. “Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap.Cell Motil Cytoskeleton, vol. 18, no. 1, 1991, pp. 55–62. Pubmed, doi:10.1002/cm.970180106.
Voter WA, O’Brien ET, Erickson HP. Dilution-induced disassembly of microtubules: relation to dynamic instability and the GTP cap. Cell Motil Cytoskeleton. 1991;18(1):55–62.
Journal cover image

Published In

Cell Motil Cytoskeleton

DOI

ISSN

0886-1544

Publication Date

1991

Volume

18

Issue

1

Start / End Page

55 / 62

Location

United States

Related Subject Headings

  • Tubulin
  • Solutions
  • Microtubules
  • Kinetics
  • Guanosine Triphosphate
  • Developmental Biology
  • Animals
  • 3101 Biochemistry and cell biology
  • 1103 Clinical Sciences
  • 0601 Biochemistry and Cell Biology