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PMID:20940262

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Citation

Schittenhelm, RB, Althoff, F, Heidmann, S and Lehner, CF (2010) Detrimental incorporation of excess Cenp-A/Cid and Cenp-C into Drosophila centromeres is prevented by limiting amounts of the bridging factor Cal1. J. Cell. Sci. 123:3768-79

Abstract

Propagation of centromere identity during cell cycle progression in higher eukaryotes depends critically on the faithful incorporation of a centromere-specific histone H3 variant encoded by CENPA in humans and cid in Drosophila. Cenp-A/Cid is required for the recruitment of Cenp-C, another conserved centromere protein. With yeast three-hybrid experiments, we demonstrate that the essential Drosophila centromere protein Cal1 can link Cenp-A/Cid and Cenp-C. Cenp-A/Cid and Cenp-C interact with the N- and C-terminal domains of Cal1, respectively. These Cal1 domains are sufficient for centromere localization and function, but only when linked together. Using quantitative in vivo imaging to determine protein copy numbers at centromeres and kinetochores, we demonstrate that centromeric Cal1 levels are far lower than those of Cenp-A/Cid, Cenp-C and other conserved kinetochore components, which scale well with the number of kinetochore microtubules when comparing Drosophila with budding yeast. Rather than providing a stoichiometric link within the mitotic kinetochore, Cal1 limits centromeric deposition of Cenp-A/Cid and Cenp-C during exit from mitosis. We demonstrate that the low amount of endogenous Cal1 prevents centromere expansion and mitotic kinetochore failure when Cenp-A/Cid and Cenp-C are present in excess.

Links

PubMed Online version:10.1242/jcs.067934

Keywords

Animals; Cells, Cultured; Centromere/metabolism; Chromosomal Proteins, Non-Histone/metabolism; DNA-Binding Proteins/metabolism; Drosophila Proteins/metabolism; Drosophila melanogaster/genetics; Histones/metabolism; Humans; Kinetochores/metabolism; Microtubules/metabolism; Mitosis/genetics; Two-Hybrid System Techniques; Yeasts/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DROME:Q8T9G0

involved_in

GO:0000278: mitotic cell cycle

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q8T9G0

located_in

GO:0000775: chromosome, centromeric region

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q8T9G0

enables

GO:0030674: protein-macromolecule adaptor activity

ECO:0000353: physical interaction evidence used in manual assertion

FB:FBgn0040477,FB:FBgn0266916

F

Seeded From UniProt

complete

DROME:Q8T9G0

located_in

GO:0005730: nucleolus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q8T9G0

involved_in

GO:0071459: protein localization to chromosome, centromeric region

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q8T9G0

involved_in

GO:0034501: protein localization to kinetochore

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q9VEN2

involved_in

GO:0000278: mitotic cell cycle

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q9VEN2

located_in

GO:0000775: chromosome, centromeric region

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q9VEN2

enables

GO:0030674: protein-macromolecule adaptor activity

ECO:0000353: physical interaction evidence used in manual assertion

FB:FBgn0040477,FB:FBgn0266916

F

Seeded From UniProt

complete

DROME:Q9VEN2

located_in

GO:0005730: nucleolus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q9VEN2

involved_in

GO:0071459: protein localization to chromosome, centromeric region

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q9VEN2

involved_in

GO:0034501: protein localization to kinetochore

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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