EMSA Analysis of DNA Binding By Rgg Proteins.

Bio Protoc

Medicinal Chemstry and Pharmacognosy, University of Illionois at Chicago, Chicago, USA.

Published: August 2013

In bacteria, interaction of various proteins with DNA is essential for the regulation of specific target gene expression. Electrophoretic mobility shift assay (EMSA) is an approach allowing for the visualization of these protein-DNA interactions. Rgg proteins comprise a family of transcriptional regulators widespread among the Firmicutes. Some of these proteins function independently to regulate target gene expression, while others have now been demonstrated to function as effectors of cell-to-cell communication, having regulatory activities that are modulated via direct interaction with small signaling peptides. EMSA analysis can be used to assess DNA binding of either type of Rgg protein. EMSA analysis of Rgg protein activity has facilitated confirmation of regulatory targets, identification of precise DNA binding sites via DNA probe mutagenesis, and characterization of the mechanism by which some cognate signaling peptides modulate Rgg protein function ( interruption of DNA-binding in some cases).

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4943666PMC
http://dx.doi.org/10.21769/bioprotoc.838DOI Listing

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