Next Article in Journal
Predicting miRNA-Disease Associations by Incorporating Projections in Low-Dimensional Space and Local Topological Information
Previous Article in Journal
Transcript Abundance Patterns of 9- and 13-Lipoxygenase Subfamily Gene Members in Response to Abiotic Stresses (Heat, Cold, Drought or Salt) in Tomato (Solanum lycopersicum L.) Highlights Member-Specific Dynamics Relevant to Each Stress
Open AccessCommunication

A Simple Method to Detect the Inhibition of Transcription Factor-DNA Binding Due to Protein–Protein Interactions In Vivo

State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and Technology, Guangxi University, Nanning 530005, China
*
Author to whom correspondence should be addressed.
Genes 2019, 10(9), 684; https://doi.org/10.3390/genes10090684
Received: 18 July 2019 / Revised: 3 September 2019 / Accepted: 4 September 2019 / Published: 6 September 2019
(This article belongs to the Section Technologies and Resources for Genetics)
Binding of transcription factors (TFs) to cis-regulatory elements (DNA) could modulate the expression of downstream genes, while interactions between TFs and other proteins might inhibit them binding to DNA. Nowadays, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) approaches are usually employed to detect the inhibitory effect. However, EMSA might not reflect the inhibitory effect in vivo. ChIP requires preparation of specific antibody or stable genetic transformation and complicated experimental steps, making it laborious and time-consuming. Here, based on the yeast one-hybrid (Y1H) system, we present a simple method to detect the inhibition of TF–DNA binding due to protein–protein interactions in vivo. When interactions between TFs and other proteins inhibit TFs binding to DNA, the reporter (Aureobasidin A resistance) gene is not activated, thereby inhibiting yeast growth on media containing the AbA antibiotic. Two examples were tested with the newly developed method to demonstrate its feasibility. In conclusion, this method provides an alternative strategy for detecting the inhibition of DNA-binding of TFs due to their interactions with other proteins in vivo. View Full-Text
Keywords: protein–DNA binding; protein–protein interactions; yeast one-hybrid; EMSA; ChIP protein–DNA binding; protein–protein interactions; yeast one-hybrid; EMSA; ChIP
Show Figures

Figure 1

MDPI and ACS Style

Yang, G.; Chao, D.; Ming, Z.; Xia, J. A Simple Method to Detect the Inhibition of Transcription Factor-DNA Binding Due to Protein–Protein Interactions In Vivo. Genes 2019, 10, 684.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map

1
Back to TopTop