Deenah Osman
The Effectors and Sensory Sites of Formaldehyde-Responsive Regulator FrmR and Metal-Sensing Variant
Osman, Deenah; Piergentili, Cecilia; Chen, Junjun; Sayer, Lucy; Usón, Isabel; Huggins, Thomas; Robinson, Nigel; Pohl, Ehmke
Authors
Cecilia Piergentili
Junjun Chen
Lucy Sayer
Isabel Usón
Thomas Huggins
Professor Nigel Robinson nigel.robinson@durham.ac.uk
Emeritus Professor
Professor Ehmke Pohl ehmke.pohl@durham.ac.uk
Interim Director
Abstract
The DUF156 family of DNA-binding, transcriptional-regulators include metal-sensors which respond to cobalt and/or nickel (RcnR, InrS) or copper (CsoR), plus CstR which responds to persulfide, and formaldehyde-responsive FrmR. Unexpectedly, the allosteric mechanism of FrmR from Salmonella enterica serovar Typhimurium is triggered by metals in vitro and variant FrmRE64H gains responsiveness to Zn(II) and cobalt in vivo. Here we establish that the allosteric mechanism of FrmR is triggered directly by formaldehyde in vitro. Sensitivity to formaldehyde requires a cysteine (Cys35 in FrmR) conserved in all DUF156 proteins. A crystal structure of metal- and formaldehyde-sensing FrmRE64H reveals that an FrmR-specific amino-terminal Pro2 is proximal to Cys35 and these residues form the deduced formaldehyde-sensing site. Evidence is presented which implies that residues spatially close to the conserved cysteine tune the sensitivities of DUF156 proteins above or below critical thresholds for different effectors, generating the semblance of specificity within cells. Relative to FrmR, RcnR is less responsive to formaldehyde in vitro and RcnR does not sense formaldehyde in vivo, but reciprocal mutations FrmRP2S and RcnRS2P respectively impair or enhance formaldehyde-reactivity in vitro. Formaldehyde-detoxification by FrmA requires S-(hydroxymethyl)glutathione, yet glutathione inhibits formaldehyde detection by FrmR in vivo and in vitro. Quantifying the number of FrmR molecules per cell and modelling formaldehyde modification as a function of [formaldehyde], demonstrates that FrmR-reactivity is optimised such that FrmR is modified, and frmRA de-repressed, at lower [formaldehyde] than required to generate S-(hydroxymethyl)glutathione. Expression of FrmA is thereby coordinated with the accumulation of its substrate.
Citation
Osman, D., Piergentili, C., Chen, J., Sayer, L., Usón, I., Huggins, T., …Pohl, E. (2016). The Effectors and Sensory Sites of Formaldehyde-Responsive Regulator FrmR and Metal-Sensing Variant. Journal of Biological Chemistry, 291(37), 19502-19516. https://doi.org/10.1074/jbc.m116.745174
Journal Article Type | Article |
---|---|
Acceptance Date | Jul 29, 2016 |
Online Publication Date | Jul 29, 2016 |
Publication Date | Sep 1, 2016 |
Deposit Date | Aug 1, 2016 |
Publicly Available Date | Aug 1, 2016 |
Journal | Journal of Biological Chemistry |
Print ISSN | 0021-9258 |
Electronic ISSN | 1083-351X |
Publisher | American Society for Biochemistry and Molecular Biology |
Peer Reviewed | Peer Reviewed |
Volume | 291 |
Issue | 37 |
Pages | 19502-19516 |
DOI | https://doi.org/10.1074/jbc.m116.745174 |
Public URL | https://durham-repository.worktribe.com/output/1407024 |
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Copyright Statement
This final published version is available under a Creative Commons CC-BY license.
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