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Molecular and Cellular Biology, October 2005, p. 8971-8984, Vol. 25, No. 20
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.20.8971-8984.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Condensin I Subunit Barren/CAP-H Is Essential for the Structural Integrity of Centromeric Heterochromatin during Mitosis{dagger}

Raquel A. Oliveira,1,2 Paula A. Coelho,1 and Claudio E. Sunkel1,3*

Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal,1 PDBEB, Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal,2 ICBAS, Universidade do Porto, Largo do Prof. Abel Salazar, 4000 Porto, Portugal3

Received 6 May 2005/ Returned for modification 22 June 2005/ Accepted 11 July 2005

During cell division, chromatin undergoes structural changes essential to ensure faithful segregation of the genome. Condensins, abundant components of mitotic chromosomes, are known to form two different complexes, condensins I and II. To further examine the role of condensin I in chromosome structure and in particular in centromere organization, we depleted from S2 cells the Drosophila CAP-H homologue Barren, a subunit exclusively associated with condensin I. In the absence of Barren/CAP-H the condensin core subunits DmSMC4/2 still associate with chromatin, while the other condensin I non-structural maintenance of chromosomes family proteins do not. Immunofluorescence and in vivo analysis of Barren/CAP-H-depleted cells showed that mitotic chromosomes are able to condense but fail to resolve sister chromatids. Additionally, Barren/CAP-H-depleted cells show chromosome congression defects that do not appear to be due to abnormal kinetochore-microtubule interaction. Instead, the centromeric and pericentromeric heterochromatin of Barren/CAP-H-depleted chromosomes shows structural problems. After bipolar attachment, the centromeric heterochromatin organized in the absence of Barren/CAP-H cannot withstand the forces exerted by the mitotic spindle and undergoes irreversible distortion. Taken together, our data suggest that the condensin I complex is required not only to promote sister chromatid resolution but also to maintain the structural integrity of centromeric heterochromatin during mitosis.

* Corresponding author. Mailing address: Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal. Phone: 351-22-6074900. Fax: 351-22-6099157. E-mail: cesunkel{at}ibmc.up.pt.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org.

Molecular and Cellular Biology, October 2005, p. 8971-8984, Vol. 25, No. 20
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.20.8971-8984.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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