RRC ID 50621
Author Sugino H, Usui T, Shimada T, Nakano M, Ogasawara H, Ishihama A, Hirata A.
Title A structural sketch of RcdA, a transcription factor controlling the master regulator of biofilm formation.
Journal FEBS Lett
Abstract RcdA is a regulator of curlin subunit gene D, the master regulator of biofilm formation in Escherichia coli. Here, we determined the X-ray structure of RcdA at 2.55 Å resolution. RcdA consists of an N-terminal DNA-binding domain (DBD) containing a helix-turn-helix (HTH) motif and a C-terminal dimerization domain, and forms a homodimer in crystals. A computational docking model of the RcdA-DNA complex allowed prediction of the candidate residues responsible for DNA binding. Our structure-guided mutagenesis, in combination with gel shift assay, atomic force microscopic observation, and reporter assay, indicate that R32 in α2 of the HTH motif plays an essential role in the recognition and binding of target DNA while T46 in α3 influences the mode of oligomerization. These results provide insights into the DNA-binding mode of RcdA.
Volume 591(13)
Pages 2019-2031
Published 2017-7-1
DOI 10.1002/1873-3468.12713
PMID 28608551
MeSH Biofilms / growth & development* DNA, Bacterial / chemistry DNA, Bacterial / metabolism DNA-Binding Proteins / chemistry* DNA-Binding Proteins / metabolism* Escherichia coli / metabolism Escherichia coli / physiology Escherichia coli Proteins / chemistry* Escherichia coli Proteins / metabolism* Models, Molecular Nucleic Acid Conformation Protein Conformation, alpha-Helical Transcription Factors / chemistry* Transcription Factors / metabolism*
IF 3.057
Times Cited 3
Resource
Prokaryotes E. coli ME9780(Escherichia coli K12 W3110 type-A)