Co(II)-detection does not follow Kco(II) gradient: channelling in Co(II)-sensing

Patterson, C.J.; Pernil, R.; Dainty, S.J.; Chakrabarti, B.; Henry, C.E.; Money, V.A; Foster, A.W.; Robinson, N.J.

Abstract

The MerR-like transcriptional activator CoaR detects surplus Co(II) to regulate Co(II) efflux in a cyanobacterium. This organism also has cytosolic metal-sensors from three further families represented by Zn(II)-sensors ZiaR and Zur plus Ni(II)-sensor InrS. Here we discover by competition with Fura-2 that CoaR has KCo(II) weaker than 7 × 10−8 M, which is weaker than ZiaR, Zur and InrS (KCo(II) = 6.94 ± 1.3 × 10−10 M; 4.56 ± 0.16 × 10−10 M; and 7.69 ± 1.1 × 10−9 M respectively). KCo(II) for CoaR is also weak in the CoaR–DNA adduct. Further, Co(II) promotes DNA-dissociation by ZiaR and DNA-association by Zur in vitro in a manner analogous to Zn(II), as monitored by fluorescence anisotropy. After 48 h exposure to maximum non-inhibitory [Co(II)], CoaR responds in vivo yet the two Zn(II)-sensors do not, despite their tighter KCo(II) and despite Co(II) triggering allostery in ZiaR and Zur in vitro. These data imply that the two Zn(II) sensors fail to respond because they fail to gain access to Co(II) under these conditions in vivo. Several lines of evidence suggest that CoaR is membrane associated via a domain with sequence similarity to precorrin isomerase, an enzyme of vitamin B12 biosynthesis. Moreover, site directed mutagenesis reveals that transcriptional activation requires CoaR residues that are predicted to form hydrogen bonds to a tetrapyrrole. The Co(II)-requiring vitamin B12 biosynthetic pathway is also membrane associated suggesting putative mechanisms by which Co(II)-containing tetrapyrroles and/or Co(II) ions are channelled to CoaR.

Citation

Patterson, C., Pernil, R., Dainty, S., Chakrabarti, B., Henry, C., Money, V., …Robinson, N. (2013). Co(II)-detection does not follow Kco(II) gradient: channelling in Co(II)-sensing. Metallomics, 5(4), 352-362. https://doi.org/10.1039/c3mt20241k

• Journal Article Type: Article
• Publication Date: Apr 1, 2013
• Deposit Date: Jan 14, 2015
• Publicly Available Date: Jan 20, 2015
• Journal: Metallomics
• Print ISSN: 1756-5901
• Electronic ISSN: 1756-591X
• Publisher: Oxford University Press
• Peer Reviewed: Peer Reviewed
• Volume: 5
• Issue: 4
• Pages: 352-362
• DOI: https://doi.org/10.1039/c3mt20241k
• Public URL: https://durham-repository.worktribe.com/output/1447215