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PMID: 17333235 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Chromosoma ·Vol. 116 ·No. 4 ·2007-08-00 ·Pages 385-402

Schittenhelm RB, Heeger S, Althoff F, Walter A, Heidmann S, Mechtler K, Lehner CF

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.

MeSH Terms
Animals Animals, Genetically Modified Centromere/chemistry,ultrastructure Centromere Protein A Chromosomal Proteins, Non-Histone/analysis Chromosomes/chemistry,ultrastructure DNA-Binding Proteins/analysis Drosophila Drosophila Proteins/analysis,physiology Histones/analysis Kinetochores/chemistry,ultrastructure Microtubule-Associated Proteins/analysis Mitosis Nuclear Proteins/analysis,physiology Spindle Apparatus/chemistry
Chemicals
Centromere Protein A Chromosomal Proteins, Non-Histone Cid protein, Drosophila DNA-Binding Proteins Drosophila Proteins Histones Microtubule-Associated Proteins Nuclear Proteins centromere protein C
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Schittenhelm Ralf B
Department of Genetics, University of Bayreuth, Universitaetsstr. 30, 95447 Bayreuth, Germany.
Heeger Sebastian
Althoff Friederike
Walter Anne
Heidmann Stefan
Mechtler Karl
Lehner Christian F
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Article Info
Journal
Chromosoma
Abbr.
Chromosoma
ISSN
0009-5915
DOI
10.1007/s00412-007-0103-y
Published
2007-08-00
Epub
2007-00-01
Pages
385-402
Language
English
Region
Austria
NLM ID
2985138R
PMCID
PMC1950589
Subset
IM
External Links
PubMed source DOI: 10.1007/s00412-007-0103-y PMC Full Text View journal details
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