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Title: Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans

Abstract

Four highly similar genes (W08E12.2, W08E12.3, W08E12.4 and W08E12.5) which are consecutively aligned on chromosome IV of the C. elegans genome are predicted to code for small (120–141aa) yet cysteine rich (18–19 cysteines) proteins. Cloning and sequencing of the genomic regions of the isoforms confirmed the presence and order of all genes. The generation of transgenic worms strains with an integrated single copy or extrachromosomal multi-copy PW08E12.3;W08E12.4::GFP uncovered that W08E12.3 and W08E12.4 are constitutively expressed in the pharynx and significantly induced in worms exposed to 100 μM Zn. Knockdown by RNAi did not have a marked consequence on reproductive performance nor was a Zn-dependent effect on nematode growth observed. However, RNAi of these genes led to an accumulation of Zn in the intestinal cells. W08E12.3 was recombinantly expressed in E. coli and the purified protein was shown to be able to bind up to 6.5 Zn molecules at neutral pH. Zn-binding was acid-labile and the apo protein was observed at pH < 4.3. In conclusion, this characterization suggests W08E12.2, W08E12.3, W08E12.4 and W08E12.5 belong to a family of putative Metalloproteins which, akin to metallothioneins, may play an important role in Zn-sensing, homeostasis and/or detoxification.

Authors:
 [1];  [2];  [1];  [1];  [3]; ORCiD logo [2]; ORCiD logo [1]
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  1. King's College London, School of Population Health & Environmental Sciences, Faculty of Life Sciences & Medicine, London, UK
  2. University of Warwick, Department of Chemistry, Coventry, UK
  3. Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, USA
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1482410
Alternate Identifier(s):
OSTI ID: 1490674
Grant/Contract Number:  
AC02-76SF00515; BB/E025064; BB/E025099
Resource Type:
Published Article
Journal Name:
Metallomics
Additional Journal Information:
Journal Name: Metallomics Journal Volume: 10 Journal Issue: 12; Journal ID: ISSN 1756-5901
Publisher:
Oxford University Press
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chaudhuri, Poulami, Imam, Hasan Tanvir, Essig, Yona, Krasauskas, Jovaras, Webb, Samuel M., Blindauer, Claudia A., and Stürzenbaum, Stephen R. Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans. United Kingdom: N. p., 2018. Web. doi:10.1039/C8MT00169C.
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Chaudhuri, Poulami, Imam, Hasan Tanvir, Essig, Yona, Krasauskas, Jovaras, Webb, Samuel M., Blindauer, Claudia A., & Stürzenbaum, Stephen R. Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans. United Kingdom. https://doi.org/10.1039/C8MT00169C
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Chaudhuri, Poulami, Imam, Hasan Tanvir, Essig, Yona, Krasauskas, Jovaras, Webb, Samuel M., Blindauer, Claudia A., and Stürzenbaum, Stephen R. Wed . "Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans". United Kingdom. https://doi.org/10.1039/C8MT00169C.
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@article{osti_1482410,
title = {Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans},
author = {Chaudhuri, Poulami and Imam, Hasan Tanvir and Essig, Yona and Krasauskas, Jovaras and Webb, Samuel M. and Blindauer, Claudia A. and Stürzenbaum, Stephen R.},
abstractNote = {Four highly similar genes (W08E12.2, W08E12.3, W08E12.4 and W08E12.5) which are consecutively aligned on chromosome IV of the C. elegans genome are predicted to code for small (120–141aa) yet cysteine rich (18–19 cysteines) proteins. Cloning and sequencing of the genomic regions of the isoforms confirmed the presence and order of all genes. The generation of transgenic worms strains with an integrated single copy or extrachromosomal multi-copy PW08E12.3;W08E12.4::GFP uncovered that W08E12.3 and W08E12.4 are constitutively expressed in the pharynx and significantly induced in worms exposed to 100 μM Zn. Knockdown by RNAi did not have a marked consequence on reproductive performance nor was a Zn-dependent effect on nematode growth observed. However, RNAi of these genes led to an accumulation of Zn in the intestinal cells. W08E12.3 was recombinantly expressed in E. coli and the purified protein was shown to be able to bind up to 6.5 Zn molecules at neutral pH. Zn-binding was acid-labile and the apo protein was observed at pH < 4.3. In conclusion, this characterization suggests W08E12.2, W08E12.3, W08E12.4 and W08E12.5 belong to a family of putative Metalloproteins which, akin to metallothioneins, may play an important role in Zn-sensing, homeostasis and/or detoxification.},
doi = {10.1039/C8MT00169C},
journal = {Metallomics},
number = 12,
volume = 10,
place = {United Kingdom},
year = {Wed Dec 12 00:00:00 EST 2018},
month = {Wed Dec 12 00:00:00 EST 2018}
}
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Journal Article:
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Publisher's Version of Record
https://doi.org/10.1039/C8MT00169C
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Figures / Tables:

Fig. 1 Fig. 1: (A) Schematic diagram of a section of cosmid W08E12 which is part of chromosome IV depicting the location of W08E12.2, W08E12.3, W08E12.4 and W08E12.5. Note the coding sequences of W08E12.4 and W08E12.5 (red boxes), and the first 500 bp of the W08E12.3 and W08E12.4 promoters (red dashed lines)more » are identical. (B) ClustalW alignment of amino acid sequences of W08E12.2, W08E12.3, W08E12.4 and W08E12.5. Identical residues are shown in black and cysteines are highlighted in red. Note: the high level of sequence identity between W08E12.3, W08E12.4 and W08E12.5, with W08E12.2 being shorter and lacking the N terminal cysteine.« less
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Works referenced in this record:

Caenorhabditis elegans: An Emerging Model in Biomedical and Environmental Toxicology
journal, June 2008

  • Leung, Maxwell C. K.; Williams, Phillip L.; Benedetto, Alexandre
  • Toxicological Sciences, Vol. 106, Issue 1
  • DOI: 10.1093/toxsci/kfn121

Use of keratin fiber for separation of heavy metals from water
journal, October 2004

  • Kar, P.; Misra, M.
  • Journal of Chemical Technology & Biotechnology, Vol. 79, Issue 11
  • DOI: 10.1002/jctb.1132

Structure and properties of keratin/PEO blend nanofibres
journal, August 2008

Caenorhabditis elegans expresses a functional phytochelatin synthase: C. elegans phytochelatin synthase
journal, July 2001

Caenorhabditis elegans Metallothioneins Protect against Toxicity Induced by Depleted Uranium
journal, July 2009

  • Jiang, George C. -T.; Hughes, Sam; Stürzenbaum, Stephen R.
  • Toxicological Sciences, Vol. 111, Issue 2
  • DOI: 10.1093/toxsci/kfp161

A comparative approach using ecotoxicological methods from single-species bioassays to model ecosystems : Comparing bioassays from single-species tests to model ecosystems
journal, July 2016

  • Haegerbaeumer, Arne; Höss, Sebastian; Ristau, Kai
  • Environmental Toxicology and Chemistry, Vol. 35, Issue 12
  • DOI: 10.1002/etc.3482

The Metabolomic Responses of Caenorhabditis elegans to Cadmium Are Largely Independent of Metallothionein Status, but Dominated by Changes in Cystathionine and Phytochelatins
journal, July 2009

  • Hughes, Samantha L.; Bundy, Jacob G.; Want, Elizabeth J.
  • Journal of Proteome Research, Vol. 8, Issue 7
  • DOI: 10.1021/pr9001806

Phytochelatin Synthase Genes from Arabidopsis and the Yeast Schizosaccharomyces pombe
journal, June 1999

  • Ha, Suk-Bong; Smith, Aaron P.; Howden, Ross
  • The Plant Cell, Vol. 11, Issue 6
  • DOI: 10.1105/tpc.11.6.1153

Sublethal Toxicity Endpoints of Heavy Metals to the Nematode Caenorhabditis elegans
journal, January 2016

The Possible Role of Cytokeratin 8 in Cadmium-Induced Adaptation and Carcinogenesis
journal, March 2007

C.elegans Metallothioneins: New Insights into the Phenotypic Effects of Cadmium Toxicosis
journal, August 2004

  • Swain, S. C.; Keusekotten, K.; Baumeister, R.
  • Journal of Molecular Biology, Vol. 341, Issue 4
  • DOI: 10.1016/j.jmb.2004.06.050

Phytochelatins and Their Roles in Heavy Metal Detoxification
journal, July 2000

Copy-number changes in evolution: rates, fitness effects and adaptive significance
journal, January 2013

The Cation Diffusion Facilitator Gene cdf-2 Mediates Zinc Metabolism in Caenorhabditis elegans
journal, May 2009

Elemental and Chemically Specific X-ray Fluorescence Imaging of Biological Systems
journal, January 2014

  • Pushie, M. Jake; Pickering, Ingrid J.; Korbas, Malgorzata
  • Chemical Reviews, Vol. 114, Issue 17
  • DOI: 10.1021/cr4007297

Deletion of Phytochelatin Synthase Modulates the Metal Accumulation Pattern of Cadmium Exposed C. elegans
journal, February 2016

  • Essig, Yona; Webb, Samuel; Stürzenbaum, Stephen
  • International Journal of Molecular Sciences, Vol. 17, Issue 2
  • DOI: 10.3390/ijms17020257

Zn- and Cd-Metallothionein Recombinant Species from the Most Diverse Phyla May Contain Sulfide (S2−) Ligands
journal, July 2005

  • Capdevila, Mercè; Domènech, Jordi; Pagani, Ayelén
  • Angewandte Chemie International Edition, Vol. 44, Issue 29
  • DOI: 10.1002/anie.200501183

Genome-scale analysis of in vivo spatiotemporal promoter activity in Caenorhabditis elegans
journal, May 2007

  • Dupuy, Denis; Bertin, Nicolas; Hidalgo, César A.
  • Nature Biotechnology, Vol. 25, Issue 6
  • DOI: 10.1038/nbt1305

Expression of tandem gene duplicates is often greater than twofold
journal, May 2016

  • Loehlin, David W.; Carroll, Sean B.
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 21
  • DOI: 10.1073/pnas.1605886113

Hair Keratin Associated Proteins: Characterization of a Second High Sulfur KAP Gene Domain on Human Chromosome 2111In fond memory of Dr Peter Steinert.
journal, January 2004

Transcriptional Profile of Aging in C. elegans
journal, September 2002

Using the Nematode Caenorhabditis Elegans To Predict Mammalian Acute Lethality To Metallic Salts
journal, October 1988

A New Pathway for Heavy Metal Detoxification in Animals: PHYTOCHELATIN SYNTHASE IS REQUIRED FOR CADMIUM TOLERANCE IN
journal, April 2001

  • Vatamaniuk, Olena K.; Bucher, Elizabeth A.; Ward, James T.
  • Journal of Biological Chemistry, Vol. 276, Issue 24
  • DOI: 10.1074/jbc.C100152200

Expression profiling of five different xenobiotics using a Caenorhabditis elegans whole genome microarray
journal, October 2005

Tools for metal ion sorting: in vitro evidence for partitioning of zinc and cadmium in C. elegans metallothionein isoforms
journal, January 2011

  • Leszczyszyn, Oksana I.; Zeitoun-Ghandour, Sukaina; Stürzenbaum, Stephen R.
  • Chem. Commun., Vol. 47, Issue 1
  • DOI: 10.1039/C0CC02188A

MgATP-Dependent Transport of Phytochelatins Across the Tonoplast of Oat Roots
journal, April 1995

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