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Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes

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 Drosophil...

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Autores principales: Schittenhelm, Ralf B., Heeger, Sebastian, Althoff, Friederike, Walter, Anne, Heidmann, Stefan, Mechtler, Karl, Lehner, Christian F.
Formato: Texto
Lenguaje:English
Publicado: Springer-Verlag 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1950589/
https://www.ncbi.nlm.nih.gov/pubmed/17333235
http://dx.doi.org/10.1007/s00412-007-0103-y
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author Schittenhelm, Ralf B.
Heeger, Sebastian
Althoff, Friederike
Walter, Anne
Heidmann, Stefan
Mechtler, Karl
Lehner, Christian F.
author_facet Schittenhelm, Ralf B.
Heeger, Sebastian
Althoff, Friederike
Walter, Anne
Heidmann, Stefan
Mechtler, Karl
Lehner, Christian F.
author_sort Schittenhelm, Ralf B.
collection PubMed
description 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.
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spelling pubmed-19505892007-08-22 Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes Schittenhelm, Ralf B. Heeger, Sebastian Althoff, Friederike Walter, Anne Heidmann, Stefan Mechtler, Karl Lehner, Christian F. Chromosoma Research Article 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. Springer-Verlag 2007-03-01 2007-08 /pmc/articles/PMC1950589/ /pubmed/17333235 http://dx.doi.org/10.1007/s00412-007-0103-y Text en © Springer-Verlag 2007
spellingShingle Research Article
Schittenhelm, Ralf B.
Heeger, Sebastian
Althoff, Friederike
Walter, Anne
Heidmann, Stefan
Mechtler, Karl
Lehner, Christian F.
Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title_full Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title_fullStr Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title_full_unstemmed Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title_short Spatial organization of a ubiquitous eukaryotic kinetochore protein network in Drosophila chromosomes
title_sort spatial organization of a ubiquitous eukaryotic kinetochore protein network in drosophila chromosomes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1950589/
https://www.ncbi.nlm.nih.gov/pubmed/17333235
http://dx.doi.org/10.1007/s00412-007-0103-y
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