scholarly journals Cytoplasmic CopZ-Like Protein and Periplasmic Rusticyanin and AcoP Proteins as Possible Copper Resistance Determinants in Acidithiobacillus ferrooxidans ATCC 23270

2015 ◽  
Vol 82 (4) ◽  
pp. 1015-1022 ◽  
Author(s):  
Claudio A. Navarro ◽  
Diego von Bernath ◽  
Cristóbal Martínez-Bussenius ◽  
Rodrigo A. Castillo ◽  
Carlos A. Jerez
Keyword(s):  
Acidithiobacillus Ferrooxidans ◽  
Binding Proteins ◽  
Metal Tolerance ◽  
Metal Resistance ◽  
Copper Resistance ◽  
Copper Binding ◽  
Content Type ◽  
Resistance Determinants ◽  
High Level ◽  
Copper Binding Proteins

ABSTRACTAcidophilic organisms, such asAcidithiobacillus ferrooxidans, possess high-level resistance to copper and other metals.A. ferrooxidanscontains canonical copper resistance determinants present in other bacteria, such as CopA ATPases and RND efflux pumps, but these components do not entirely explain its high metal tolerance. The aim of this study was to find other possible copper resistance determinants in this bacterium. Transcriptional expression ofA. ferrooxidansgenes coding for a cytoplasmic CopZ-like copper-binding chaperone and the periplasmic copper-binding proteins rusticyanin and AcoP, which form part of an iron-oxidizing supercomplex, was found to increase when the microorganism was grown in the presence of copper. All of these proteins conferred more resistance to copper when expressed heterologously in a copper-sensitiveEscherichia colistrain. This effect was absent when site-directed-mutation mutants of these proteins with altered copper-binding sites were used in this metal sensitivity assay. These results strongly suggest that the three copper-binding proteins analyzed here are copper resistance determinants in this extremophile and contribute to the high-level metal resistance of this industrially important biomining bacterium.

scholarly journals Multiple Megaplasmids Confer Extremely High Levels of Metal Tolerance in Alteromonas Strains

2019 ◽  
Vol 86 (3) ◽  
Author(s):  
Kathleen D. Cusick ◽  
Shawn W. Polson ◽  
Gabriel Duran ◽  
Russell T. Hill
Keyword(s):  
Gene Transfer ◽  
Pseudomonas Syringae ◽  
Molecular Mechanisms ◽  
Metal Tolerance ◽  
Bioinformatic Analysis ◽  
Metal Resistance ◽  
Copper Resistance ◽  
The Other ◽  
Copper Tolerance ◽  
Content Type

ABSTRACT Alteromonas is a widely distributed genus of marine Gammaproteobacteria, with representatives shown to be key players in diverse processes, including biogeochemical cycling and biofouling of marine substrata. While Alteromonas spp. are early colonizers of copper-based antifouling paints on marine vessels, their mechanism of tolerance is poorly understood. PacBio whole-genome sequencing of Alteromonas macleodii strains CUKW and KCC02, isolated from Cu/Ni alloy test coupons submerged in oligotrophic coastal waters, indicated the presence of multiple megaplasmids (ca. 200 kb) in both. A pulsed-field gel electrophoresis method was developed and used to confirm the presence of multiple megaplasmids in these two strains; it was then used to screen additional Alteromonas strains for which little to no sequencing data exist. Plasmids were not detected in any of the other strains. Bioinformatic analysis of the CUKW and KCC02 plasmids identified numerous genes associated with metal resistance. Copper resistance orthologs from both the Escherichia coli Cue and Cus and Pseudomonas syringae Cop systems were present, at times as multiple copies. Metal growth assays in the presence of copper, cobalt, manganese, and zinc performed with 10 Alteromonas strains demonstrated the ability of CUKW and KCC02 to grow at metal concentrations inhibitory to all the other strains tested. This study reports multiple megaplasmids in Alteromonas strains. Bioinformatic analysis of the CUKW and KCC02 plasmids indicate that they harbor elements of the Tra system conjugation apparatus, although their type of mobility remains to be experimentally verified. IMPORTANCE Copper is commonly used as an antifouling agent on ship hulls. Alteromonas spp. are early colonizers of copper-based antifouling paint, but their mechanism of tolerance is poorly understood. Sequencing of A. macleodii strains isolated from copper test materials for marine ships indicated the presence of multiple megaplasmids. Plasmids serve as key vectors in horizontal gene transfer and confer traits such as metal resistance, detoxification, ecological interaction, and antibiotic resistance. Bioinformatic analysis identified many metal resistance genes and genes associated with mobility. Understanding the molecular mechanisms and capacity for gene transfer within marine biofilms provides a platform for the development of novel antifouling solutions targeting genes involved in copper tolerance and biofilm formation.

Rapid induction of copper-binding proteins in the gills of metal exposed mussels

1980 ◽  
Vol 67 (2) ◽  
pp. 215-218 ◽  
Author(s):  
A. Viarengo ◽  
M. Pertica ◽  
G. Mancinelli ◽  
G. Zanicchi ◽  
M. Orunesu
Keyword(s):  
Binding Proteins ◽  
Copper Binding ◽  
Rapid Induction ◽  
Copper Binding Proteins

Zinc- and copper-binding proteins in the plasma of winter flounder (Pseudopleuronectes americanus)

1980 ◽  
Vol 58 (4) ◽  
pp. 609-613 ◽  
Author(s):  
P. E. Fletcher ◽  
G. L. Fletcher
Keyword(s):  
Molecular Weight ◽  
Binding Proteins ◽  
Binding Protein ◽  
Gel Filtration ◽  
Protein S ◽  
Egg Yolk ◽  
Winter Flounder ◽  
Zinc Binding ◽  
Copper Binding ◽  
Copper Binding Proteins

Zinc- and copper-binding proteins were isolated from the plasma of winter flounder using gel filtration chromatography. A single copper-binding protein fraction of molecular weight 170 000 was isolated from the plasma of both sexes.In male and female flounder over 95% of the plasma zinc was associated with a zinc-binding protein(s) with a molecular weight of 76 000. In male flounder the remaining zinc appeared to be bound to a protein(s) of molecular weight 186 000. In female flounder the remaining 5% of the zinc was associated with two zinc-binding fractions with apparent molecular weights of 186 000 and 340 000 – 370 000.Extracts of plasma vitellogenin and egg yolk proteins revealed significant quantities of zinc and copper. It is hypothesized that the female specific zinc-binding protein (340 000 – 370 000) was vitellogenin.

Stability and Folding of Copper-Binding Proteins

2010 ◽  
pp. 61-80
Author(s):  
Irina Pozdnyakova ◽  
Pernilla Wittung-Stafshede
Keyword(s):  
Binding Proteins ◽  
Copper Binding ◽  
Copper Binding Proteins

High molecular weight zinc- and copper-binding proteins in the liver of the rat; Effect of centrifugation

1987 ◽  
Vol 135 (2) ◽  
pp. 129-132
Author(s):  
J.B. Dielhof, ◽  
J.G. Van Der Enden, ◽  
TH.M.W. Stolk ◽  
C.J.A. Van Den Hamer,
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Binding Proteins ◽  
Copper Binding ◽  
Copper Binding Proteins

scholarly journals Transcriptomes of the Extremely Thermoacidophilic Archaeon Metallosphaera sedula Exposed to Metal “Shock” Reveal Generic and Specific Metal Responses

2016 ◽  
Vol 82 (15) ◽  
pp. 4613-4627 ◽  
Author(s):  
Garrett H. Wheaton ◽  
Arpan Mukherjee ◽  
Robert M. Kelly
Keyword(s):  
Metal Toxicity ◽  
Cellular Response ◽  
Sulfur Metabolism ◽  
Metal Resistance ◽  
Open Reading Frames ◽  
Copper Resistance ◽  
Natural Environments ◽  
Cluster Assembly ◽  
Content Type ◽  
Metallosphaera Sedula

ABSTRACTThe extremely thermoacidophilic archaeonMetallosphaera sedulamobilizes metals by novel membrane-associated oxidase clusters and, consequently, requires metal resistance strategies. This issue was examined by “shocking”M. sedulawith representative metals (Co2+, Cu2+, Ni2+, UO22+, Zn2+) at inhibitory and subinhibitory levels. Collectively, one-quarter of the genome (554 open reading frames [ORFs]) responded to inhibitory levels, and two-thirds (354) of the ORFs were responsive to a single metal. Cu2+(259 ORFs, 106 Cu2+-specific ORFs) and Zn2+(262 ORFs, 131 Zn2+-specific ORFs) triggered the largest responses, followed by UO22+(187 ORFs, 91 UO22+-specific ORFs), Ni2+(93 ORFs, 25 Ni2+-specific ORFs), and Co2+(61 ORFs, 1 Co2+-specific ORF). While one-third of the metal-responsive ORFs are annotated as encoding hypothetical proteins, metal challenge also impacted ORFs responsible for identifiable processes related to the cell cycle, DNA repair, and oxidative stress. Surprisingly, there were only 30 ORFs that responded to at least four metals, and 10 of these responded to all five metals. This core transcriptome indicated induction of Fe-S cluster assembly (Msed_1656-Msed_1657), tungsten/molybdenum transport (Msed_1780-Msed_1781), and decreased central metabolism. Not surprisingly, a metal-translocating P-type ATPase (Msed_0490) associated with a copper resistance system (Cop) was upregulated in response to Cu2+(6-fold) but also in response to UO22+(4-fold) and Zn2+(9-fold). Cu2+challenge uniquely induced assimilatory sulfur metabolism for cysteine biosynthesis, suggesting a role for this amino acid in Cu2+resistance or issues in sulfur metabolism. The results indicate thatM. sedulaemploys a range of physiological and biochemical responses to metal challenge, many of which are specific to a single metal and involve proteins with yet unassigned or definitive functions.IMPORTANCEThe mechanisms by which extremely thermoacidophilic archaea resist and are negatively impacted by metals encountered in their natural environments are important to understand so that technologies such as bioleaching, which leverage microbially based conversion of insoluble metal sulfides to soluble species, can be improved. Transcriptomic analysis of the cellular response to metal challenge provided both global and specific insights into how these novel microorganisms negotiate metal toxicity in natural and technological settings. As genetics tools are further developed and implemented for extreme thermoacidophiles, information about metal toxicity and resistance can be leveraged to create metabolically engineered strains with improved bioleaching characteristics.

Isolation of copper-binding proteins from activated sludge culture

2001 ◽  
Vol 44 (2-3) ◽  
pp. 453-459 ◽  
Author(s):  
K. Fukushi ◽  
S. Kato ◽  
T. Antsuki ◽  
T. Omura
Keyword(s):  
Molecular Weight ◽  
Activated Sludge ◽  
Metal Uptake ◽  
Binding Proteins ◽  
Growth Media ◽  
Copper Binding ◽  
Molecular Weights ◽  
Binding Capability ◽  
Copper Binding Proteins

Six copper-binding microbial proteins were isolated from activated sludge cultures grown on media containing copper at various concentrations. Molecular weights among isolated proteins were ranged from 1.3k to 174k dalton. Isolated proteins were compared for their copper binding capabilities. Proteins isolated from cultures grown in the presence of copper in the growth media exhibited higher copper binding capabilities than those isolated from the culture grown in the absence of copper. The highest metal uptake of 61.23 (mol copper/mol protein) was observed by a protein isolated from a culture grown with copper at a concentration of 0.25 mM. This isolated protein (CBP2) had a molecular weight of 24k dalton. Other protein exhibited copper binding capability of 4.8-32.5 (mol copper/mol protein).

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