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J. Biol. Chem., Vol. 280, Issue 16, 16437-16445, April 22, 2005

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Beyond the Xi

MacroH2A CHROMATIN DISTRIBUTION AND POST-TRANSLATIONAL MODIFICATION IN AN AVIAN SYSTEM^*

D. Wade Abbott, Brian P. Chadwick, Anita A. Thambirajah, and Juan Ausió¶

From the Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8W 3P6, Canada and the Department of Molecular Genetics and Microbiology, Institute for Genome Sciences and Policy, Duke University Medical Center, Durham, North Carolina 27710

MacroH2A (mH2A) is a histone variant that is enriched in the inactivated X-chromosomes of mammalian females. To characterize the role of this protein in other nuclear processes we isolated chromatin particles from chicken liver, a vertebrate system that does not undergo X-inactivation. Chromatin digestion and fractionation studies determined that mH2A is evenly distributed at several levels of chromatin structure and stabilizes the nucleosome core particle in solution. However, at the level of the chromatosome, selective salt precipitation showed the existence of a mutually exclusive relationship between mH2A and H1, which may reveal functional redundancy between these proteins. Two-dimensional gel electrophoresis demonstrated the presence of one major population of mH2A containing nucleosomes, which may become ADP-ribosylated. This report provides new clues into how mH2A distribution and a previously unidentified post-translational modification may help regulate the repression of autosomal chromatin.

Received for publication, January 6, 2005 , and in revised form, February 14, 2005.

^* This work was supported by Canadian Institutes of Health Research Grant MOP-57718 (to J. A.) and a Postgraduate Scholarship Natural Sciences and Engineering Research Council of Canada Doctoral Scholarship (to D. W. A.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom all correspondence should be addressed. Tel.: 250-721-8863; Fax: 250-721-8855; E-mail: jausio{at}uvic.ca.

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