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A chromatin remodelling complex that loads cohesin onto human chromosomes

Abstract

Nucleosomal DNA is arranged in a higher-order structure that presents a barrier to most cellular processes involving protein DNA interactions1. The cellular machinery involved in sister chromatid cohesion, the cohesin complex, also requires access to the nucleosomal DNA to perform its function in chromosome segregation2,3,4,5,6,7,8,9,10. The machineries that provide this accessibility are termed chromatin remodelling factors11. Here, we report the isolation of a human ISWI (SNF2h)-containing chromatin remodelling complex that encompasses components of the cohesin and NuRD complexes. We show that the hRAD21 subunit of the cohesin complex directly interacts with the ATPase subunit SNF2h. Mapping of hRAD21, SNF2h and Mi2 binding sites by chromatin immunoprecipitation experiments reveals the specific association of these three proteins with human DNA elements containing Alu sequences. We find a correlation between modification of histone tails and association of the SNF2h/cohesin complex with chromatin. Moreover, we show that the association of the cohesin complex with chromatin can be regulated by the state of DNA methylation. Finally, we present evidence pointing to a role for the ATPase activity of SNF2h in the loading of hRAD21 on chromatin.

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Figure 1: Isolation of an SNF2h complex containing cohesin.
Figure 2: Affinity-purification of the SNF2h/cohesin complex.
Figure 3: Association of SNF2h/Mi2/cohesin complex with human DNA containing Alu repeats.
Figure 4: Roles of DNA methylation and chromatin remodelling on cohesin loading.

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Acknowledgements

We thank W. Wang and G. Mandel for the gift of Mi2 and REST antibodies, respectively. We also thank T. Nagase at the Kazusa DNA Research Institute in Japan for providing the hRAD21 cDNA. We also thank H. C. Gregson and A. R. Ball, Jr for analysis of Rad21 and SNF2h interactions. This work was supported by a grant from NIH to R.S. and in part by a March of Dimes Basil O'Conner Scholarship and the NIH to K.Y. K.Y. is a Scholar of the Leukemia & Lymphoma Society. M.-A.H. was supported by a postdoctoral fellowship from Association pour la Recherche sur le Cancer (FRANCE). D.A.B. is a recipient of an NIH training grant.

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Correspondence to Ramin Shiekhattar.

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Hakimi, MA., Bochar, D., Schmiesing, J. et al. A chromatin remodelling complex that loads cohesin onto human chromosomes. Nature 418, 994–998 (2002). https://doi.org/10.1038/nature01024

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