Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter

Lana C. Ly; Yang Yang; Fudong Li; Merlin Crossley; Yunyu Shi; Kate G.R. Quinlan

STAR Protocols (Sep 2022)

Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter

Lana C. Ly,
Yang Yang,
Fudong Li,
Merlin Crossley,
Yunyu Shi,
Kate G.R. Quinlan

Affiliations

Lana C. Ly: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia; Corresponding author
Yang Yang: School of Life Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230026, China; Ministry of Education Key Laboratory for Membraneless Organelles and Cellular Dynamics, University of Science and Technology of China, Hefei, China
Fudong Li: Ministry of Education Key Laboratory for Membraneless Organelles and Cellular Dynamics, University of Science and Technology of China, Hefei, China
Merlin Crossley: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia
Yunyu Shi: School of Life Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230026, China; Ministry of Education Key Laboratory for Membraneless Organelles and Cellular Dynamics, University of Science and Technology of China, Hefei, China; Corresponding author
Kate G.R. Quinlan: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia; Corresponding author

Journal volume & issue: Vol. 3, no. 3
p. 101598

Abstract

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Summary: Here, we describe protocols to interrogate the binding of the zinc fingers of the transcription factor ZBTB7A to the fetal γ-globin (HBG) promoter. We detail the steps for performing electrophoretic mobility shift assays (EMSAs), X-ray crystallography, and isothermal titration calorimetry (ITC) to explore this interaction. These techniques could readily be applied to the structural studies of other zinc finger transcription factors and cognate DNA sequences.For complete details on the use and execution of this protocol, please refer to Yang et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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