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Structure of Centrosomes and Chromosomes Through IVEM Tomography

Published online by Cambridge University Press:  02 July 2020

D.A. Agard
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA, 94143-0448
Vincent Guenbaut
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA, 94143-0448
Michelle Moritz
Affiliation:
Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA, 94143-0448.
M.B. Braunfeld
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA, 94143-0448
GuoFung Zhang
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA, 94143-0448
John W. Sedat
Affiliation:
Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA, 94143-0448.
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Extract

Centrosome structure: The centrosome is the major microtubule-organizing center of animal cells - it initiates spindle formation at the onset of mitosis and plays an active role in chromosome segregation. At the most basic level, microtubule (MT) formation is regulated at least in part, through the functional availability of nucleation sites on the centrosome. Functional centrosomes can be isolated from Drosophila embryos. When visualized with IVEM Tomography (IVEM-T) ring complexes are visualized throughout the peri-centriolar material (PCM) (1). γ-tubulin has been implicated in MT nucleation. Through immuno-IVEM-T we have shown that the ring structures within the PCM, contain y-tubulin and further that the only place γ-tubulin is found is at the minus (nucleating) end of MTs (2). This strongly implicates these γ-tubulin containing ring structures in MT polymerization, until our studies there was no direct proof that γ-tubulin was located at the site of MT nucleation. Furthermore Zheng et al have been able to isolate similar ring structures that are soluble from both Xenopus and Drosophila embryos (3).

Type
Innovative Approaches to 3-D Structure/Function Determination for Cells and Organelles
Copyright
Copyright © Microscopy Society of America 1997

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References

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