Cytosolic Ni(II) sensor in cyanobacterium: nickel detection follows nickel affinity across four families of metal sensors.

The Journal of Biological Chemistry
Andrew W FosterNigel J Robinson

Abstract

Efflux of surplus Ni(II) across the outer and inner membranes of Synechocystis PCC 6803 is mediated by the Nrs system under the control of a sensor of periplasmic Ni(II), NrsS. Here, we show that the product of ORF sll0176, which encodes a CsoR/RcnR-like protein now designated InrS (for internal nickel-responsive sensor), represses nrsD (NrsD is deduced to efflux Ni(II) across the inner membrane) from a cryptic promoter between the final two ORFs in the nrs operon. Transcripts initiated from the newly identified nrsD promoter accumulate in response to nickel or cobalt but not copper, and recombinant InrS forms specific, Ni(II)-inhibited complexes with the nrsD promoter region. Metal-dependent difference spectra of Ni(II)- and Cu(I)-InrS are similar to Cu(I)-sensing CsoR and dissimilar to Ni(II)/Co(II)-sensing RcnR, consistent with factors beyond the primary coordination sphere switching metal selectivity. Competition with chelators mag-fura-2, nitrilotriacetic acid, EDTA, and EGTA estimate K(D) Ni(II) for the tightest site of InrS as 2.05 (±1.5) × 10(-14) m, and weaker K(D) Ni(II) for the cells' metal sensors of other types: Zn(II) co-repressor Zur, Co(II) activator CoaR, and Zn(II) derepressor ZiaR. Ni(II) transfer to InrS occ...Continue Reading

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Citations

Sep 14, 2012·Biochemistry·Khadine A HigginsMichael J Maroney
Feb 20, 2013·Metallomics : Integrated Biometal Science·Carl J PattersonNigel J Robinson
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Nov 11, 2021·Nucleic Acids Research·Joseph N FakhouryDavid P Giedroc

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Methods Mentioned

BETA
PCR
electrophoretic mobility shift
Fluorescence
size exclusion chromatography

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