Studying RNA Polymerase II Promoter-Proximal Pausing by In Vitro Immobilized Template and Transcription Assays

Heeyoun Bunch

doi:10.1007/978-1-0716-3342-7_2

Studying RNA Polymerase II Promoter-Proximal Pausing by In Vitro Immobilized Template and Transcription Assays

Methods Mol Biol. 2023:2693:13-24. doi: 10.1007/978-1-0716-3342-7_2.

Author

Heeyoun Bunch¹

Affiliation

¹ Department of Applied Biosciences, Kyungpook National University, Daegu, Republic of Korea.

PMID: 37540423
DOI: 10.1007/978-1-0716-3342-7_2

Abstract

The immobilized template assay is a versatile biochemical method for studying protein-nucleic acid interactions. Using this method, immobilized nucleic acid-associated or specific proteins can be identified and quantified by techniques such as mass spectrometry and immunoblotting. Here, a modified immobilized template assay combined with in vitro transcription assay to study the function of transcription factors and transcriptional activities at the human heat shock protein 70 (HSP70) gene is described. Notably, this method can be applied to study other important genes and transcription factors in vitro.

Keywords: Gene regulation; Immobilized template assay; In vitro transcription; Transcription factors; Transcriptional regulation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

HSP70 Heat-Shock Proteins / metabolism
Humans
Nucleic Acids*
Promoter Regions, Genetic
RNA Polymerase II* / metabolism
Transcription Factors / metabolism
Transcription, Genetic

Substances

RNA Polymerase II
Transcription Factors
HSP70 Heat-Shock Proteins
Nucleic Acids