Abstract
The male-specific lethal (MSL) complex upregulates the single male X chromosome to achieve dosage compensation in Drosophila melanogaster. We have proposed that MSL recognition of specific entry sites on the X is followed by local targeting of active genes marked by histone H3 trimethylation (H3K36me3). Here we analyze the role of the MSL3 chromodomain in the second targeting step. Using ChIP-chip analysis, we find that MSL3 chromodomain mutants retain binding to chromatin entry sites but show a clear disruption in the full pattern of MSL targeting in vivo, consistent with a loss of spreading. Furthermore, when compared to wild type, chromodomain mutants lack preferential affinity for nucleosomes containing H3K36me3 in vitro. Our results support a model in which activating complexes, similarly to their silencing counterparts, use the nucleosomal binding specificity of their respective chromodomains to spread from initiation sites to flanking chromatin.
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Acknowledgements
Special thanks to A.A. Alekseyenko (Harvard-Partners Center for Genetics and Genomics) for important advice, discussions and protocols, S. Khorasanizadeh for discussions and to members of the Kuroda laboratory for critical comments on the manuscript, H. Oh (currently at Rutgers University), for a key fly stock, and J. Racine and A. Sarovschii for technical assistance and the Taplin Biological Mass Spectrometry Facility (Harvard Medical School) for peptide identification. This work was supported by the US National Institutes of Health (NIH; GM45744 to M.I.K. and GM67825 to P.J.P.). B.L. and J.L.W. were supported by GM47867 from the NIH and by funding from the Stowers Institute for Medical Research.
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T.H.S. performed the mutagenesis, transgenic complementation, immunoprecipitations, ChIP-chip, purification of recombinant MSL3 and polytene-chromosome analyses; S.P. performed all bioinformatics analyses; B.L. performed the in vitro nucleosome binding assays; J.L.W., P.J.P. and M.I.K. supervised the analyses; T.H.S. and M.I.K. prepared the manuscript in consultation with all coauthors.
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Sural, T., Peng, S., Li, B. et al. The MSL3 chromodomain directs a key targeting step for dosage compensation of the Drosophila melanogaster X chromosome. Nat Struct Mol Biol 15, 1318–1325 (2008). https://doi.org/10.1038/nsmb.1520
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DOI: https://doi.org/10.1038/nsmb.1520
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