Copper binding and reactivity at the histidine brace motif: insights from mutational analysis of the Pseudomonas fluorescens copper chaperone CopC

FEBS Letters ◽  
2021 ◽  
Author(s):  
Johan Ø. Ipsen ◽  
Cristina Hernández‐Rollán ◽  
Sebastian J. Muderspach ◽  
Søren Brander ◽  
Andreas B. Bertelsen ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Mutational Analysis ◽  
Copper Chaperone ◽  
Copper Binding

Copper chaperone Atox1 interacts with the metal-binding domain of Wilson's disease protein in cisplatin detoxification

2013 ◽  
Vol 454 (1) ◽  
pp. 147-156 ◽  
Author(s):  
Nataliya V. Dolgova ◽  
Sergiy Nokhrin ◽  
Corey H. Yu ◽  
Graham N. George ◽  
Oleg Y. Dmitriev
Keyword(s):  
Metal Binding ◽  
Wilson’S Disease ◽  
Wilson's Disease ◽  
Binding Domain ◽  
Binding Motif ◽  
Copper Chaperone ◽  
Copper Binding ◽  
Copper Transporters ◽  
Metal Binding Domain ◽  
Disease Protein

Human copper transporters ATP7B (Wilson's disease protein) and ATP7A (Menkes' disease protein) have been implicated in tumour resistance to cisplatin, a widely used anticancer drug. Cisplatin binds to the copper-binding sites in the N-terminal domain of ATP7B, and this binding may be an essential step of cisplatin detoxification involving copper ATPases. In the present study, we demonstrate that cisplatin and a related platinum drug carboplatin produce the same adduct following reaction with MBD2 [metal-binding domain (repeat) 2], where platinum is bound to the side chains of the cysteine residues in the CxxC copper-binding motif. This suggests the same mechanism for detoxification of both drugs by ATP7B. Platinum can also be transferred to MBD2 from copper chaperone Atox1, which was shown previously to bind cisplatin. Binding of the free cisplatin and reaction with the cisplatin-loaded Atox1 produce the same protein-bound platinum intermediate. Transfer of platinum along the copper-transport pathways in the cell may serve as a mechanism of drug delivery to its target in the cell nucleus, and explain tumour-cell resistance to cisplatin associated with the overexpression of copper transporters ATP7B and ATP7A.

scholarly journals A putative nuclear copper chaperone promotes plant immunity in Arabidopsis

2020 ◽  
Vol 71 (20) ◽  
pp. 6684-6696 ◽  
Author(s):  
Long-Xiang Chai ◽  
Kai Dong ◽  
Song-Yu Liu ◽  
Zhen Zhang ◽  
Xiao-Peng Zhang ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Plant Immunity ◽  
Copper Chaperone ◽  
Copper Binding ◽  
Nuclear Speckles ◽  
Defense Gene Expression ◽  
Copper Chaperones ◽  
Plant Nucleus

Abstract Copper is essential for many metabolic processes but must be sequestrated by copper chaperones. It is well known that plant copper chaperones regulate various physiological processes. However, the functions of copper chaperones in the plant nucleus remain largely unknown. Here, we identified a putative copper chaperone induced by pathogens (CCP) in Arabidopsis thaliana. CCP harbors a classical MXCXXC copper-binding site (CBS) at its N-terminus and a nuclear localization signal (NLS) at its C-terminus. CCP mainly formed nuclear speckles in the plant nucleus, which requires the NLS and CBS domains. Overexpression of CCP induced PR1 expression and enhanced resistance against Pseudomonas syringae pv. tomato DC3000 compared with Col-0 plants. Conversely, two CRISPR/Cas9-mediated ccp mutants were impaired in plant immunity. Further biochemical analyses revealed that CCP interacted with the transcription factor TGA2 in vivo and in vitro. Moreover, CCP recruits TGA2 to the PR1 promoter sequences in vivo, which induces defense gene expression and plant immunity. Collectively, our results have identified a putative nuclear copper chaperone required for plant immunity and provided evidence for a potential function of copper in the salicylic pathway.

scholarly journals Mutational Analysis of the Saccharomyces cerevisiae Cytochrome c Oxidase Assembly Protein Cox11p

2006 ◽  
Vol 5 (3) ◽  
pp. 568-578 ◽  
Author(s):  
Graham S. Banting ◽  
D. Moira Glerum
Keyword(s):  
Hydrogen Peroxide ◽  
Cytochrome Oxidase ◽  
Solution Structure ◽  
Sinorhizobium Meliloti ◽  
Mutational Analysis ◽  
Intermembrane Space ◽  
Copper Binding ◽  
Inner Mitochondrial Membrane ◽  
The Matrix

ABSTRACT Cox11p is an integral protein of the inner mitochondrial membrane that is essential for cytochrome c oxidase assembly. The bulk of the protein is located in the intermembrane space and displays high levels of evolutionary conservation. We have analyzed a collection of site-directed and random cox11 mutants in an effort to further define essential portions of the molecule. Of the alleles studied, more than half had no apparent effect on Cox11p function. Among the respiration deficiency-encoding alleles, we identified three distinct phenotypes, which included a set of mutants with a misassembled or partially assembled cytochrome oxidase, as indicated by a blue-shifted cytochrome aa 3 peak. In addition to the shifted spectral signal, these mutants also display a specific reduction in the levels of subunit 1 (Cox1p). Two of these mutations are likely to occlude a surface pocket behind the copper-binding domain in Cox11p, based on analogy with the Sinorhizobium meliloti Cox11 solution structure, thereby suggesting that this pocket is crucial for Cox11p function. Sequential deletions of the matrix portion of Cox11p suggest that this domain is not functional beyond the residues involved in mitochondrial targeting and membrane insertion. In addition, our studies indicate that Δcox11, like Δsco1, displays a specific hypersensitivity to hydrogen peroxide. Our studies provide the first evidence at the level of the cytochrome oxidase holoenzyme that Cox1p is the in vivo target for Cox11p and suggest that Cox11p may also have a role in the response to hydrogen peroxide exposure.

Copper binding modulates the platination of human copper chaperone Atox1 by antitumor trans-platinum complexes

Metallomics ◽  
2014 ◽  
Vol 6 (3) ◽  
pp. 491-497 ◽  
Author(s):  
Zhaoyong Xi ◽  
Wei Guo ◽  
Changlin Tian ◽  
Fuyi Wang ◽  
Yangzhong Liu
Keyword(s):  
Platinum Complexes ◽  
Copper Chaperone ◽  
Copper Binding

Cu(i) coordination enhances the reactivity of Atox1 towards antitumor-active trans-platinum complexes and promotes platinum transfer from the protein to dithiothreitol.

Copper binding promotes the interaction of cisplatin with human copper chaperone Atox1

2013 ◽  
Vol 49 (95) ◽  
pp. 11197 ◽  
Author(s):  
Zhaoyong Xi ◽  
Wei Guo ◽  
Changlin Tian ◽  
Fuyi Wang ◽  
Yangzhong Liu
Keyword(s):  
Copper Chaperone ◽  
Copper Binding

scholarly journals GacS Sensor Domains Pertinent to the Regulation of Exoproduct Formation and to the Biocontrol Potential of Pseudomonas fluorescens CHA0

2003 ◽  
Vol 16 (7) ◽  
pp. 634-644 ◽  
Author(s):  
Sophie Zuber ◽  
Fiona Carruthers ◽  
Christoph Keel ◽  
Alexandre Mattart ◽  
Caroline Blumer ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Mutational Analysis ◽  
Response Regulator ◽  
Single Copy ◽  
Stationary Growth Phase ◽  
Sensor Kinase ◽  
Elevated Expression ◽  
Crown And Root Rot ◽  
Cell Densities ◽  
Biocontrol Strain

In the root-colonizing biocontrol strain CHA0 of Pseudomonas fluorescens, cell density-dependent synthesis of extra-cellular, plant-beneficial secondary metabolites and enzymes is positively regulated by the GacS/GacA two-component system. Mutational analysis of the GacS sensor kinase using improved single-copy vectors showed that inactivation of each of the three conserved phosphate acceptor sites caused an exoproduct null phenotype (GacS¯), whereas deletion of the periplasmic loop domain had no significant effect on the expression of exoproduct genes. Strain CHA0 is known to synthesize a solvent-extractable extracellular signal that advances and enhances the expression of exoproduct genes during the transition from exponential to stationary growth phase when maximal exoproduct formation occurs. Mutational inactivation of either GacS or its cognate response regulator GacA abolished the strain's response to added signal. Deletion of the linker domain of the GacS sensor kinase caused signal-independent, strongly elevated expression of exoproduct genes at low cell densities. In contrast to the wild-type strain CHA0, the gacS linker mutant and a gacS null mutant were unable to protect tomato plants from crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici in a soil-less microcosm, indicating that, at least in this plant-pathogen system, there is no advantage in using a signal-independent biocontrol strain.

scholarly journals DNA Sequence and Mutational Analysis of Rhizobitoxine Biosynthesis Genes in Bradyrhizobium elkanii

2001 ◽  
Vol 67 (11) ◽  
pp. 4999-5009 ◽  
Author(s):  
Tsuyoshi Yasuta ◽  
Shin Okazaki ◽  
Hisayuki Mitsui ◽  
Ken-Ichi Yuhashi ◽  
Hiroshi Ezura ◽  
...  
Keyword(s):  
Dna Sequence ◽  
Pseudomonas Syringae ◽  
Insertional Mutagenesis ◽  
Mutational Analysis ◽  
Fatty Acid Desaturase ◽  
Random Mutagenesis ◽  
Evolutionary Significance ◽  
Broad Host Range ◽  
Copper Binding ◽  
Bradyrhizobium Elkanii

ABSTRACT We cloned and sequenced a cluster of genes involved in the biosynthesis of rhizobitoxine, a nodulation enhancer produced by Bradyrhizobium elkanii. The nucleotide sequence of the cloned 28.4-kb DNA region encompassing rtxAshowed that several open reading frames (ORFs) were located downstream of rtxA. A large-deletion mutant of B. elkanii, USDA94Δrtx::Ω1, which lacks rtxA, ORF1 (rtxC), ORF2, and ORF3, did not produce rhizobitoxine, dihydrorhizobitoxine, or serinol. The broad-host-range cosmid pLAFR1, which contains rtxAand these ORFs, complemented rhizobitoxine production in USDA94Δrtx::Ω1. Further complementation experiments involving cosmid derivatives obtained by random mutagenesis with a kanamycin cassette revealed that at least rtxAand rtxC are necessary for rhizobitoxine production. Insertional mutagenesis of the N-terminal and C-terminal regions ofrtxA indicated that rtxA is responsible for two crucial steps, serinol formation and dihydrorhizobitoxine biosynthesis. An insertional mutant of rtxC produced serinol and dihydrorhizobitoxine but no rhizobitoxine. Moreover, thertxC product was highly homologous to the fatty acid desaturase of Pseudomonas syringae and included the copper-binding signature and eight histidine residues conserved in membrane-bound desaturase. This result suggested thatrtxC encodes dihydrorhizobitoxine desaturase for the final step of rhizobitoxine production. In light of results from DNA sequence comparison, gene disruption experiments, and dihydrorhizobitoxine production from various substrates, we discuss the biosynthetic pathway of rhizobitoxine and its evolutionary significance in bradyrhizobia.

scholarly journals Selenocysteine Positional Variants Reveal Contributions to Copper Binding from Cysteine Residues in Domains 2 and 3 of Human Copper Chaperone for Superoxide Dismutase†

Biochemistry ◽  
2008 ◽  
Vol 47 (49) ◽  
pp. 13074-13083 ◽  
Author(s):  
Amanda N. Barry ◽  
Kevin M. Clark ◽  
Adenike Otoikhian ◽  
Wilfred A. van der Donk ◽  
Ninian J. Blackburn
Keyword(s):  
Superoxide Dismutase ◽  
Cysteine Residues ◽  
Copper Chaperone ◽  
Copper Binding

scholarly journals Copper-mediated dimerization of CopZ, a predicted copper chaperone from Bacillus subtilis

2002 ◽  
Vol 368 (3) ◽  
pp. 729-739 ◽  
Author(s):  
Margaret A. KIHLKEN ◽  
Andrew P. LEECH ◽  
Nick E. LE BRUN
Keyword(s):  
Bacillus Subtilis ◽  
Metal Binding ◽  
Metal Ion ◽  
Complex Nature ◽  
Binding Motif ◽  
Copper Chaperone ◽  
Ion Transfer ◽  
Copper Binding ◽  
Binding Properties ◽  
In The Wild

Understanding the metal-binding properties and solution states of metallo-chaperones is a key step in understanding how they function in metal ion transfer. Using spectroscopic, bioanalytical and biochemical methods, we have investigated the copper-binding properties and association states of the putative copper chaperone of Bacillus subtilis, CopZ, and a variant of the protein lacking the two cysteine residues of the MXCXXC copper-binding motif. We show that copper-free CopZ exists as a monomer, but that addition of copper(I) causes the protein to associate into homodimers. The nature of the copper(I)—CopZ complex is dependent on the level of copper loading, and we report the detection of three distinct forms, containing 0.5, 1.0 and 1.5 copper(I) ions per protein. The presence of excess dithiothreitol has a significant effect on copper(I) binding to CopZ, such that, in its presence, copper(I)—CopZ occurs mainly as a monomer species. Data for copper binding to the double-cysteine variant of CopZ are consistent with an essential role for these residues in tight copper binding in the wild-type protein. We conclude that the complex nature of copper(I) binding to CopZ may underpin mechanisms of protein-to-protein copper(I) transfer.

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