MTT2, a copper-inducible metallothionein gene from Tetrahymena thermophila

Boldrin, Francesco; Santovito, Gianfranco; Formigari, Alessia; Bisharyan, Yelena; Cassidy-Hanley, Donna; Clark, Theodore G.; Piccinni, E.

doi:10.1016/j.cbpc.2007.10.002

Cited by 31 publications

(32 citation statements)

References 37 publications

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“…(29,30,36,37) Both subfamilies differ in the induction patterns by heavy metals (Cd/Zn or Cu) and the pattern of Cys residue clustering. (29,30,31,34,37,38) The majority of Tetrahymena MT genes have been isolated as DNA complementary to RNA (cDNA) molecules, and from their alignment to the corresponding genomic DNA sequences, the absence of introns in all these MT genes has been revealed.…”

Section: Tetrahymena Mts Classificationmentioning

confidence: 99%

“…Indeed, both TtheMTT1 or TtheMTT2 promoters have been already used for overexpression of homolog or heterolog genes in T. thermophila, and are thus important biotechnological tools for expressing eukaryotic proteins, some of which are difficult to express in prokaryotic cells. (34,36,89,90) Likewise, to detect heavy metals in polluted environmental samples using a Tetrahymena whole-cell biosensor, it is possible to construct promoter-reporter gene fusions using TtheMTT1 or TtheMTT5 promoters and the green fluorescent protein or luciferase as reporter genes.…”

Section: Biotechnological Applicationsmentioning

confidence: 99%

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From heavy metal‐binders to biosensors: Ciliate metallothioneins discussed

2009

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Metallothioneins (MTs) are ubiquitous proteins with the capacity to bind heavy metal ions (mainly Cd, Zn or Cu), and they have been found in animals, plants, eukaryotic and prokaryotic micro-organisms. We have carried out a comparative analysis of ciliate MTs (Tetrahymena species) to well-known MTs from other organisms, discussing their exclusive features, such as the presence of aromatic amino acid residues and almost exclusive cysteine clusters (CCC) present in cadmium-binding metallothioneins (CdMTs), higher heavy metal-MT stoichiometry values, and a strictly conserved modular-submodular structure. Based on this last feature and an extensive gene duplication, we propose a possible model for the evolutionary history of T. thermophila MTs. We also suggest possible functions for these MTs from consideration of their differential gene expressions and discuss the potential use of these proteins and/or their gene promoters for designing molecular or whole-cell biosensors for a fast detection of heavy metals in diverse polluted ecosystems.

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Section: Tetrahymena Mts Classificationmentioning

confidence: 99%

Section: Biotechnological Applicationsmentioning

confidence: 99%

From heavy metal‐binders to biosensors: Ciliate metallothioneins discussed

2009

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“…As said above, Zn can directly interact with these sequences by binding to MTF-1 (Boldrin et al, 2008). Cd can be bound by Zn-MTs, with consequent release of Zn which can activate the MTF-1, as demonstrated by Günther et al (2012) in Drosophila melanogaster.…”

Section: Mresmentioning

confidence: 99%

Characterization and metal-induced gene transcription of two new copper zinc superoxide dismutases in the solitary ascidian Ciona intestinalis

et al. 2013

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a b s t r a c tAntioxidant enzymes are known to protect living organisms against the oxidative stress risk, also induced by metals. In the present study, we describe the purification and molecular characterization of two Cu,Zn superoxide dismutases (SODs), referred to as Ci-SODa and Ci-SODb, from Ciona intestinalis, a basal chordate widely distributed in temperate shallow seawater. The putative amino acid sequences were compared with Cu,Zn SODs from other metazoans and phylogenetic analyses indicate that the two putative Ci-SODs are more related to invertebrate SODs than vertebrate ones. Both phylogenetic and preliminary homology modeling analyses suggest that Ci-SODa and Ci-SODb are extracellular and intracellular isoform, respectively. The mRNA of the two Cu,Zn SODs was localized in hemocytes and in ovarian follicular cells, as revealed by in situ hybridization. The time course of SOD mRNA levels in the presence of three different metals showed upregulation of ci-soda and inhibition of ci-sodb. Spectrophotometric analysis confirms the presence of SOD activity in Ciona tissues. Our in silico analyses of the ci-soda promoter region revealed putative consensus sequences similar to mammalian metal-responsive elements (MRE), suggesting that the transcription of these genes directly depends on metals. These data emphasize the importance of complex metal regulation of ci-soda and ci-sodb transcription, as components of an efficient detoxification pathway allowing the survival of C. intestinalis in continued, elevated presence of metals in the environment.

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“…This is typically controlled by using various metallothionein (MTT) gene promoters, which can be induced by the addition of cadmium, mercury, copper, or zinc. The MTT1 promoter, induced by cadmium chloride (CdCl 2 ) (Shang et al 2002), is the most commonly used; however, copper inducible versions also exist (Boldrin et al 2008). In addition, the Tetrahymena Hsp70-2 promoter is highly inducible by a short heat shock (Yu et al 2012).…”

Section: Exogenous Gene Expressionmentioning

confidence: 99%

Tetrahymenaas a Unicellular Model Eukaryote: Genetic and Genomic Tools

2016

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Tetrahymena thermophila is a ciliate model organism whose study has led to important discoveries and insights into both conserved and divergent biological processes. In this review, we describe the tools for the use of Tetrahymena as a model eukaryote, including an overview of its life cycle, orientation to its evolutionary roots, and methodological approaches to forward and reverse genetics. Recent genomic tools have expanded Tetrahymena's utility as a genetic model system. With the unique advantages that Tetrahymena provide, we argue that it will continue to be a model organism of choice.

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MTT2, a copper-inducible metallothionein gene from Tetrahymena thermophila

Cited by 31 publications

References 37 publications

From heavy metal‐binders to biosensors: Ciliate metallothioneins discussed

From heavy metal‐binders to biosensors: Ciliate metallothioneins discussed

Characterization and metal-induced gene transcription of two new copper zinc superoxide dismutases in the solitary ascidian Ciona intestinalis

Tetrahymenaas a Unicellular Model Eukaryote: Genetic and Genomic Tools

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