Hamperl, Stephan and Brown, Christopher R. and Garea, Ana Villar and Perez-Fernandez, Jorge and Bruckmann, Astrid and Huber, Katharina and Wittner, Manuel and Babl, Virginia and Stoeckl, Ulrike and Deutzmann, Rainer and Boeger, Hinrich and Tschochner, Herbert and Milkereit, Philipp and Griesenbeck, Joachim (2014) Compositional and structural analysis of selected chromosomal domains from Saccharomyces cerevisiae. NUCLEIC ACIDS RESEARCH, 42 (1): e2. ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
Chromatin is the template for replication and transcription in the eukaryotic nucleus, which needs to be defined in composition and structure before these processes can be fully understood. We report an isolation protocol for the targeted purification of specific genomic regions in their native chromatin context from Saccharomyces cerevisiae. Subdomains of the multicopy ribosomal DNA locus containing transcription units of RNA polymerases I, II or III or an autonomous replication sequence were independently purified in sufficient amounts and purity to analyze protein composition and histone modifications by mass spectrometry. We present and discuss the proteomic data sets obtained for chromatin in different functional states. The native chromatin was further amenable to electron microscopy analysis yielding information about nucleosome occupancy and positioning at the single-molecule level. We also provide evidence that chromatin from virtually every single copy genomic locus of interest can be purified and analyzed by this technique.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RNA-POLYMERASE-I; MASS-SPECTROMETRIC DATA; RIBOSOMAL DNA LOCUS; CHROMATIN-STRUCTURE; PROMOTER NUCLEOSOMES; TRANSCRIPTION UNITS; GLOBAL ANALYSIS; SILENCED LOCI; PHO5 PROMOTER; RDNA REPEATS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Dr. Joachim Griesenbeck |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 29 Nov 2019 12:01 |
| Last Modified: | 29 Nov 2019 12:01 |
| URI: | https://pred.uni-regensburg.de/id/eprint/11050 |
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