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Nucleic Acids Research Advance Access originally published online on November 7, 2006
Nucleic Acids Research 2006 34(21):6247-6255; doi:10.1093/nar/gkl902
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Nucleic Acids Research, 2006, Vol. 34, No. 21 6247-6255
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Proteolysis restricts localization of CID, the centromere-specific histone H3 variant of Drosophila, to centromeres

Olga Moreno-Moreno, Mònica Torras-Llort and Fernando Azorín*

Institute of Molecular Biology of Barcelona (IBMB), CSIC, and Institute for Research in Biomedicine (IRB) Parc Científic de Barcelona, 08028 Barcelona, Spain

*To whom correspondence should be addressed at Dpto Biologia Molecular i Cel·lular, Institut de Biologia Molecular de Barcelona, CSIC, Parc Científic de Barcelona, Josep Samitier, 1-5 08028 Barcelona, Spain. Tel: 3493 403 4958; Fax: 3493 403 4979; Email: fambmc{at}ibmb.csic.es

Received July 18, 2006. Revised October 10, 2006. Accepted October 13, 2006.

Centromere identity is determined by the formation of a specialized chromatin structure containing the centromere-specific histone H3 variant CENP-A. The precise molecular mechanism(s) accounting for the specific deposition of CENP-A at centromeres are still poorly understood. Centromeric deposition of CENP-A, which is independent of DNA replication, might involve specific chromatin assembly complexes and/or specific interactions with kinetochore components. However, transiently expressed CENP-A incorporates throughout chromatin indicating that CENP-A nucleosomes can also be promiscuously deposited during DNA replication. Therefore, additional mechanisms must exist to prevent deposition of CENP-A nucleosomes during replication and/or to remove them afterwards. Here, using transient expression experiments performed in Drosophila Kc cells, we show that proteasome-mediated degradation restricts localization of Drosophila CENP-A (CID) to centromeres by eliminating mislocalized CID as well as by regulating available CID levels. Regulating available CID levels appears essential to ensure centromeric deposition of transiently expressed CID as, when expression is increased in the presence of proteasome inhibitors, newly synthesized CID mislocalizes. Mislocalization of CID affects cell cycle progression as a high percentage of cells showing mislocalized CID are reactive against {alpha}PSer10H3 antibodies, enter mitosis at a very low frequency and show strong segregation defects. However, cells showing reduced amounts of mislocalized CID show normal cell cycle progression.


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[Abstract] [Full Text] [PDF]


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