Spatial organization of a ubiquitous eukaryotic kinetochore protein network in <Emphasis Type="Italic">Drosophila</Emphasis> chromosomes |
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Authors: | Ralf B Schittenhelm Sebastian Heeger Friederike Althoff Anne Walter Stefan Heidmann Karl Mechtler Christian F Lehner |
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Institution: | (1) Department of Genetics, University of Bayreuth, Universitaetsstr. 30, 95447 Bayreuth, Germany;(2) Institute of Molecular Biotechnology GmbH, IMBA, Dr. Bohr-Gasse 3, 1030 Vienna, Austria |
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Abstract: | Chromosome segregation during meiosis and mitosis depends on the assembly of functional kinetochores within centromeric regions.
Centromeric DNA and kinetochore proteins show surprisingly little sequence conservation despite their fundamental biological
role. However, our identification in Drosophila melanogaster of the most diverged orthologs identified so far, which encode components of a kinetochore protein network including the
Ndc80 and Mis complexes, further emphasizes the notion of a shared eukaryotic kinetochore design. To determine its spatial
organization, we have analyzed by quantitative light microscopy hundreds of native chromosomes from transgenic Drosophila strains coexpressing combinations of red and green fluorescent fusion proteins, fully capable of providing the essential
wild-type functions. Thereby, Cenp-A/Cid, Cenp-C, Mis12 and the Ndc80 complex were mapped along the inter sister kinetochore
axis with a resolution below 10 nm. The C terminus of Cenp-C was found to be near but well separated from the innermost component
Cenp-A/Cid. The N terminus of Cenp-C is further out, clustered with Mis12 and the Spc25 end of the rod-like Ndc80 complex,
which is known to bind to microtubules at its other more distal Ndc80/Nuf2 end.
Ralf B. Schittenhelm and Sebastian Heeger have equal contributions. |
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