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

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 e...

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Autores principales: Ly, Lana C., Yang, Yang, Li, Fudong, Crossley, Merlin, Shi, Yunyu, Quinlan, Kate G.R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344028/
https://www.ncbi.nlm.nih.gov/pubmed/35928006
http://dx.doi.org/10.1016/j.xpro.2022.101598
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author Ly, Lana C.
Yang, Yang
Li, Fudong
Crossley, Merlin
Shi, Yunyu
Quinlan, Kate G.R.
author_facet Ly, Lana C.
Yang, Yang
Li, Fudong
Crossley, Merlin
Shi, Yunyu
Quinlan, Kate G.R.
author_sort Ly, Lana C.
collection PubMed
description 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).
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spelling pubmed-93440282022-08-03 Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter Ly, Lana C. Yang, Yang Li, Fudong Crossley, Merlin Shi, Yunyu Quinlan, Kate G.R. STAR Protoc Protocol 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). Elsevier 2022-07-31 /pmc/articles/PMC9344028/ /pubmed/35928006 http://dx.doi.org/10.1016/j.xpro.2022.101598 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Ly, Lana C.
Yang, Yang
Li, Fudong
Crossley, Merlin
Shi, Yunyu
Quinlan, Kate G.R.
Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title_full Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title_fullStr Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title_full_unstemmed Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title_short Protocols for protein-DNA binding analysis of a zinc finger transcription factor bound to its cognate promoter
title_sort protocols for protein-dna binding analysis of a zinc finger transcription factor bound to its cognate promoter
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344028/
https://www.ncbi.nlm.nih.gov/pubmed/35928006
http://dx.doi.org/10.1016/j.xpro.2022.101598
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