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

Poulami Chaudhuri, Hasan Tanvir Imam, Yona Essig, Jovaras Krasauskas, Samuel M. Webb, Claudia A. Blindauer, Stephen R. Stürzenbaum*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)
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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. 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.

Original languageEnglish
Pages (from-to)1814-1823
Number of pages10
JournalMetallomics
Volume10
Issue number12
Early online date16 Nov 2018
DOIs
Publication statusPublished - Dec 2018
Externally publishedYes

Bibliographical note

Funding Information:
This work is an extension of a joint grant funded by the Biotechnology and Biological Sciences Research Council (BBSRC grants BB/E025099 (SRS) and BB/E025064 (CAB)). Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The authors would also like to thank Phil Cunningham for his indispensable help with the bioinformatic screen. This paper is dedicated to Silvia Atrian who acted as external examiner of PC’s PhD defense.

Publisher Copyright:
© 2018 The Royal Society of Chemistry.

Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.

ASJC Scopus subject areas

  • Chemistry (miscellaneous)
  • Biophysics
  • Biomaterials
  • Biochemistry
  • Metals and Alloys

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