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Published online
doi:10.1083/jcb.200906005
The Journal of Cell Biology, Vol. 186, No. 3, 371-377
The Rockefeller University Press, 0021-9525 $30.00
© Nakanishi et al.
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Histone H2BK123 monoubiquitination is the critical determinant for H3K4 and H3K79 trimethylation by COMPASS and Dot1



Shima Nakanishi1, Jung Shin Lee1, Kathryn E. Gardner2, Jennifer M. Gardner1, Yoh-hei Takahashi1, Mahesh B. Chandrasekharan3, Zu-Wen Sun3, Mary Ann Osley4, Brian D. Strahl2, Sue L. Jaspersen1,5, and Ali Shilatifard1

1 Stowers Institute for Medical Research, Kansas City, MO 64110
2 Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC 27599
3 Department of Biochemistry, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN 37232
4 Department of Molecular Genetics and Microbiology, Albuquerque, NM 87131
5 Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160

Correspondence to Ali Shilatifard: ash{at}stowers.org

Histone H2B monoubiquitination by Rad6/Bre1 is required for the trimethylation of both histone H3K4 and H3K79 by COMPASS and Dot1 methyltransferases, respectively. The dependency of methylation at H3K4 and H3K79 on the monoubiquitination of H2BK123 was recently challenged, and extragenic mutations in the strain background used for previous studies or epitope-tagged proteins were suggested to be the sources of this discrepancy. In this study, we show that H3K4 and H3K79 methylation is solely dependent on H2B monoubiquitination regardless of any additional alteration to the H2B sequence or genome. Furthermore, we report that Y131, one of the yeast histone H2A/H2B shuffle strains widely used for the last decade in the field of chromatin and transcription biology, carries a wild-type copy of each of the HTA2 and HTB2 genes under the GAL1/10 promoter on chromosome II. Therefore, we generated the entire histone H2A and H2B alanine-scanning mutant strains in another background, which does not express wild-type histones.

© 2009 Nakanishi et al.
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

Abbreviation used in this paper: SD, synthetic dropout.


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