SAP18 Promotes Krüppel-dependent Transcriptional Repression by Enhancer-specific Histone Deacetylation*
- ‡Abteilung Molekulare Entwicklungsbiologie and ¶Bioanalytical Mass Spectrometry Group, Max-Planck-Institut für biophysikalische Chemie, Am Fassberg 11, 37077 Göttingen, Germany and the §Wadsworth Center, New York State Department of Health and Department of Biomedical Sciences, State University of New York, Albany, New York 12208
- 1 To whom correspondence should be addressed: Am Fassberg 11, 37077 Göttingen, Germany. Tel.: 49-551-201-1482/3; Fax: 49-551-201-1755; E-mail: hjaeckl{at}gwdg.de.
Abstract
Body pattern formation during early embryogenesis of Drosophila melanogaster relies on a zygotic cascade of spatially restricted transcription factor activities. The gap gene Krüppel ranks at the top level of this cascade. It encodes a C2H2 zinc finger protein that interacts directly with cis-acting stripe enhancer elements of pair rule genes, such as even skipped and hairy, at the next level of the gene hierarchy. Krüppel mediates their transcriptional repression by direct association with the corepressor Drosophila C terminus-binding protein (dCtBP). However, for some Krüppel target genes, deletion of the dCtBP-binding sites does not abolish repression, implying a dCtBP-independent mode of repression. We identified Krüppel-binding proteins by mass spectrometry and found that SAP18 can both associate with Krüppel and support Krüppel-dependent repression. Genetic interaction studies combined with pharmacological and biochemical approaches suggest a site-specific mechanism of Krüppel-dependent gene silencing. The results suggest that Krüppel tethers the SAP18 bound histone deacetylase complex 1 at distinct enhancer elements, which causes repression via histone H3 deacetylation.
Footnotes
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↵2 The abbreviations used are: HDAC, histone deacetylase; CtBP, C terminus-binding protein; ESI-MS/MS, electrospray ionization tandem mass spectrometry; HPLC, high performance liquid chromatography; HRP, horseradish peroxidase; PepSpot, peptide spot; TSA, trichostatin A; ChIP, chromatin immunoprecipitation; xChIP, cross-linked ChIP; GST, glutathione S-transferase; MALDI-TOF-MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
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↵3 A. Matyash and H. Jäckle, manuscript in preparation.
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↵* This work was supported by funds from the Max-Planck-Gesellschaft (to H. J., H. U., and A. M.) and by American Cancer Society Grant RSG-9508506-DDC and March of Dimes Grant 1-FY07-527 (to V. N. S. and S. D. H.). 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.
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The on-line version of this article (available at http://www.jbc.org) contains supplemental Tables S1 and S2 and Figs. S1 and S2.
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- Received August 8, 2008.
- Revision received November 18, 2008.
- The American Society for Biochemistry and Molecular Biology, Inc.
