Molecular genetic and biochemical characterization of a putative family of zinc metalloproteins in Caenorhabditis elegans. Issue 12 (16th November 2018)

• Record Type: Journal Article
• Authors: Chaudhuri, Poulami; Imam, Hasan Tanvir; Essig, Yona; Krasauskas, Jovaras; Webb, Samuel M.; Blindauer, Claudia A.; Stürzenbaum, Stephen R.
• Is Part Of: Metallomics. Volume 10:Issue 12(2018)
• Page Start: 1814
• Page End: 1823
• Publication Date: 2018-11-16
• Subjects: Metals -- Physiological effect -- Periodicals; 572.51
• DOI: 10.1039/c8mt00169c
• Languages: English
• ISSNs: 1756-5901

Abstract

Abstract : The first characterization of W08E12.2, W08E12.3, W08E12.4 and W08E12.5, four putative metalloproteins in C. elegans . (A) phase contrast microscopy, (B) fluorescence microscopy of PW08E12.3 ; W08E12.4 ::GFP. 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 P W08E12.3;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.