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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Detalles Bibliográficos
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
Descripción
Sumario: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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