Time-resolved analysis of transcription through chromatin

Han Wen Chang, Fu Kai Hsieh, Smita Patel, Vasily M. Studitsky

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

During transcription along nucleosomal DNA, RNA polymerase II (Pol II) pauses at multiple positions and induces formation of multiple intermediates that aid in maintaining proper chromatin structure. To describe the kinetics of this multiple-step reaction, we utilized a computational model-based approach and KinTek Explorer software to analyze the time courses. Here we describe the stepwise protocol for analysis of the kinetics of transcription through a nucleosome that provides the rate constants for each step of this complex process. We also present an example where this time-resolved approach was applied to study the mechanism of histone chaperone FACT action during Pol II transcription through a single nucleosome by comparing the rate constants derived in the presence or in the absence of FACT.

Original languageEnglish (US)
Pages (from-to)90-95
Number of pages6
JournalMethods
Volume159-160
DOIs
StatePublished - Apr 15 2019

Fingerprint

Nucleosomes
Transcription
Chromatin
Histone Chaperones
DNA Polymerase II
RNA Polymerase II
Rate constants
Kinetics
Software
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Keywords

  • Chromatin
  • Factors
  • Kinetics
  • Nucleosome
  • Transcription

Cite this

Chang, Han Wen ; Hsieh, Fu Kai ; Patel, Smita ; Studitsky, Vasily M. / Time-resolved analysis of transcription through chromatin. In: Methods. 2019 ; Vol. 159-160. pp. 90-95.
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Time-resolved analysis of transcription through chromatin. / Chang, Han Wen; Hsieh, Fu Kai; Patel, Smita; Studitsky, Vasily M.

In: Methods, Vol. 159-160, 15.04.2019, p. 90-95.

Research output: Contribution to journalArticle

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AU - Chang, Han Wen

AU - Hsieh, Fu Kai

AU - Patel, Smita

AU - Studitsky, Vasily M.

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