Profiling RNA Polymerase II Phosphorylation Genome-Wide in Fission Yeast.

Publisher: Academic Press Country of Publication: United States NLM ID: 0212271 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-7988 (Electronic) Linking ISSN: 00766879 NLM ISO Abbreviation: Methods Enzymol

Original Publication: New York, Academic Press.

RNA Polymerase II/*metabolism ; Saccharomyces cerevisiae Proteins/*metabolism ; Schizosaccharomyces/*enzymology ; Schizosaccharomyces/*metabolism; Chromatin Immunoprecipitation ; Phosphorylation ; Protein Processing, Post-Translational ; RNA Polymerase II/chemistry ; RNA Polymerase II/genetics ; Saccharomyces cerevisiae Proteins/chemistry ; Saccharomyces cerevisiae Proteins/genetics ; Schizosaccharomyces/genetics ; Transcription, Genetic/genetics

The RNA polymerase II carboxyl-terminal domain (CTD) consists of tandem repeats of consensus sequence Tyr ¹ -Ser ² -Pro ³ -Thr ⁴ -Ser ⁵ -Pro ⁶ -Ser ⁷ . Dynamic posttranslational modifications of the CTD generate a CTD code crucial for the cotranscriptional recruitment of factors that control transcription, chromatin modification, and RNA processing. Analysis of CTD phosphorylation by ChIP (Chromatin ImmunoPrecipitation) coupled with high-throughput DNA sequencing (ChIP-seq) is a powerful tool to investigate the changes in CTD phosphorylation during the transcription cycle. In this chapter, we describe a ChIP-seq protocol to profile the different CTD phospho-marks in fission yeast. Using this protocol, we have found that Tyr1P, Ser2P, and Thr4P signals are highest at gene 3' ends, whereas Ser5P is enriched across the gene bodies.
(© 2018 Elsevier Inc. All rights reserved.)

Keywords: CTD phosphorylation; ChIP-seq; Chromatin immunoprecipitation; Fission yeast; RNA polymerase II

0 (Saccharomyces cerevisiae Proteins)
EC 2.7.7.- (RNA Polymerase II)

Date Created: 20181204 Date Completed: 20190726 Latest Revision: 20190726

20221213

10.1016/bs.mie.2018.07.009

30502955