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Human chromatin platforms for the study of transcription and RNA regulation

ABSTRACT: Here we describe tools to study the step-wise assembly of protein complexes on chromatin in a highly-controlled manner using reconstituted chromatin platforms and quantitative proteomic profiling. We profile the early steps in transcriptional activation and highlight the potential for understanding the multiple ways chromatin can influence transcriptional regulation. We also describe modifications of this approach to study the activity of a long noncoding RNA to act as a dynamic scaffold for proteins to be recruited to chromatin. The reconstituted nature of the chromatin substrate offers a tune-able system, able to be trapped at specific sub-steps, to understand how chromatin interfaces with genome regulation machinery.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Saccharomyces Cerevisiae (ncbitaxon:4932)

SUBMITTER: Aaron Johnson

PROVIDER: MSV000082363 | MassIVE | Tue May 15 21:11:00 BST 2018

SECONDARY ACCESSION(S): PXD009790

REPOSITORIES: MassIVE

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Publications

SILAC-MS Profiling of Reconstituted Human Chromatin Platforms for the Study of Transcription and RNA Regulation.

Balas Maggie M MM Porman Allison M AM Hansen Kirk C KC Johnson Aaron M AM

Journal of proteome research 20180920 10

DNA packaged into chromatin is the core structure of the human genome. Nearly all eukaryotic genome regulation must interface with this genomic structure, and modification of the chromatin can influence molecular mechanisms that regulate the underlying DNA. Many processes are governed by regulated stepwise assembly mechanisms that build complex machinery on chromatin to license a specific activity such as transcription. Transcriptional activators drive the initial steps of gene expression, regul ...[more]

PMID: 30192551

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