In Vitro Assembly and Proteomic Analysis of RNA Polymerase II Complexes

Overview

Journal Methods

Specialty Biochemistry

Date 2019 Mar 8

PMID 30844430

Citations 1

Authors

Yoo Jin Joo

Scott B Ficarro

Jarrod A Marto

Stephen Buratowski

Affiliations

Soon will be listed here.

Abstract

The RNA polymerase II (RNApII) transcription cycle consists of multiple steps involving dozens of protein factors. Here we describe a useful approach to study the dynamics of initiation and early elongation, comprising an in vitro transcription system in which complexes are assembled on immobilized DNA templates and analyzed by quantitative mass spectrometry. This unbiased screening system allows quantitation of RNApII complex components on either naked DNA or chromatin templates. In addition to transcription, the system reproduces co-transcriptional mRNA capping and multiple transcription-related histone modifications. In combination with other biochemical and genetic methods, this approach can provide insights into the mechanistic details of gene expression by RNApII.

Citing Articles

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