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The Y233 gatekeeper of DmpR modulates effector‐responsive transcriptional control of σ 54 ‐RNA polymerase
Author(s) -
Seibt Henrik,
Sauer Uwe H.,
Shingler Victoria
Publication year - 2019
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1111/1462-2920.14567
Subject(s) - biology , effector , rna polymerase , polymerase , rna , transcriptional regulation , microbiology and biotechnology , genetics , gene , gene expression
Summary DmpR is the obligate transcriptional activator of genes involved in (methyl)phenol catabolism by Pseudomonas putida . DmpR belongs to the AAA + class of mechano‐transcriptional regulators that employ ATP‐hydrolysis to engage and remodel σ 54 ‐RNA polymerase to allow transcriptional initiation. Previous work has established that binding of phenolic effectors by DmpR is a prerequisite to relieve interdomain repression and allow ATP‐binding to trigger transition to its active multimeric conformation, and further that a structured interdomain linker between the effector‐ and ATP‐binding domains is involved in coupling these processes. Here, we present evidence from ATPase and in vivo and in vitro transcription assays that a tyrosine residue of the interdomain linker (Y233) serves as a gatekeeper to constrain ATP‐hydrolysis and aromatic effector‐responsive transcriptional activation by DmpR. An alanine substitution of Y233A results in both increased ATPase activity and enhanced sensitivity to aromatic effectors. We propose a model in which effector‐binding relocates Y233 to synchronize signal‐reception with multimerisation to provide physiologically appropriate sensitivity of the transcriptional response. Given that Y233 counterparts are present in many ligand‐responsive mechano‐transcriptional regulators, the model is likely to be pertinent for numerous members of this family and has implications for development of enhanced sensitivity of biosensor used to detect pollutants.