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Evidence for monomeric actin function in INO80 chromatin remodeling

Nature Structural & Molecular Biology volume 20, pages 426–432 (2013)Cite this article

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Abstract

Actin has well-established functions in the cytoplasm, but its roles in the nucleus remain poorly defined. Here, by studying the nuclear actin-containing yeast INO80 chromatin remodeling complex, we provide genetic and biochemical evidence for a role of monomeric actin in INO80 chromatin remodeling. We demonstrate that, in contrast to cytoplasmic actin, nuclear actin is present as a monomer in the INO80 complex, and its barbed end is not accessible for polymerization. We identify an actin mutation in subdomain 2 affecting in vivo nuclear functions and reducing the chromatin remodeling activity of the INO80 complex in vitro. Notably, the highly conserved subdomain 2 at the pointed end of actin contributes to the interaction of INO80 with chromatin. Our results establish an evolutionarily conserved function of nuclear actin in its monomeric form and suggest that nuclear actin can utilize a fundamentally distinct mechanism from that of cytoplasmic actin.

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Figure 1: Analysis of the actin subunit in the INO80 complex.
Figure 2: Actin contributes to INO80 chromatin remodeling.
Figure 3: Actin in the INO80 complex exists in a unique microenvironment.
Figure 4: Actin subdomain 2 is crucial for INO80 complex interaction with chromatin.
Figure 5: A model for nuclear actin in the INO80 complex.

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Acknowledgements

We thank C. Wu (National Cancer Institute), in whose laboratory we initiated early studies on actin; D. Drubin (University of California, Berkeley) for providing a collection of actin mutants; C. Boone (University of Toronto) for sharing genetic data on actin; and J. Highland, T. Wehr and J. Xiao for technical assistance. Funding to P.K. is provided by the Odyssey postdoctoral program and the Theodore N. Law Endowment for Scientific Achievement at The University of Texas MD Anderson Cancer Center. X.S. is supported by funds and grants from the US National Cancer Institute (K22CA100017), the US National Institute of General Medical Sciences (R01GM093104) and the Center for Cancer Epigenetics at MD Anderson Cancer Center.

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Author notes

  1. Prabodh Kapoor and Mingming Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, Texas, USA

    Prabodh Kapoor, Mingming Chen & Xuetong Shen

  2. Department of Biochemistry and Molecular Biology, Howard Hughes Medical Institute, Colorado State University, Fort Collins, Colorado, USA

    Duane David Winkler & Karolin Luger

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  1. Prabodh Kapoor
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Contributions

P.K., M.C. and X.S. designed the experiments. P.K. and M.C. conducted the experiments. D.D.W. and K.L. provided purified nucleosomes and performed the quantitative fluorescent nucleosome binding assays. P.K., M.C. and X.S. analyzed the data and wrote the paper.

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Correspondence to Xuetong Shen.

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The authors declare no competing financial interests.

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Kapoor, P., Chen, M., Winkler, D. et al. Evidence for monomeric actin function in INO80 chromatin remodeling. Nat Struct Mol Biol 20, 426–432 (2013). https://doi.org/10.1038/nsmb.2529

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