한빛사 논문
Abstract
Timothy W. Sikorski‡§,1,2, Yoo Jin Joo‡,1,3, Scott B. Ficarro‡§¶, Manor Askenazi‡§¶, Stephen Buratowski‡,4 and Jarrod A. Marto‡§¶,5
From the ‡Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115 and the §Department of Cancer Biology and ¶Blais Proteomics Center, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115
4 To whom correspondence may be addressed: Dept. of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Ave., Boston, MA 02115.
5 To whom correspondence may be addressed: Dept. of Cancer Biology, Dana-Farber Cancer Institute, 450 Brookline Ave., Boston, MA 02215-5450.
1 Both authors contributed equally to this work.
Abstract
In-depth characterization of RNA polymerase II preinitiation complexes remains an important and challenging goal. We used quantitative mass spectrometry to explore context-dependent Saccharomyces cerevisiae preinitiation complex formation at the HIS4 promoter reconstituted on naked and chromatinized DNA templates. The transcription activators Gal4-VP16 and Gal4-Gcn4 recruited a limited set of chromatin-related coactivator complexes, namely the chromatin remodeler Swi/Snf and histone acetyltransferases SAGA and NuA4, suggesting that transcription stimulation is mediated through these factors. Moreover, the two activators differentially recruited the coactivator complexes, consistent with specific activator-coactivator interactions. Chromatinized templates suppressed recruitment of basal transcription factors, thereby amplifying the effect of activators, compared with naked DNA templates. This system is sensitive, highly reproducible, and easily applicable to mapping the repertoire of proteins found at any promoter.
Chromatin, Histone Acetylase, Mass Spectrometry (MS), RNA Polymerase II, Transcription Coactivators, NuA4, SAGA, Swi/Snf, Transcription Activation
Footnotes
2 Supported by a National Defense Science and Engineering Graduate predoctoral fellowship.
3 Supported by a National Research Foundation of Korea postdoctoral fellowship.
* This work was supported, in whole or in part, by National Institutes of Health Grant GM46498 (to S. B.). This work was also supported by the Dana-Farber Strategic Research Initiative (to J. A. M.).
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