Molecular cloning of a cDNA and chromosomal localization of a human theta-class glutathione S-transferase gene (GSTT2) to chromosome 22

doi:10.1016/0888-7543(95)80037-M

Genomics

Volume 25, Issue 2, 20 January 1995, Pages 381-387

https://doi.org/10.1016/0888-7543(95)80037-M Get rights and content

Abstract

Until recently the Theta-class glutathione S-transferases (GSTs) were largely overlooked due to their low activity with the model substrate 1-chloro-2,4-dinitrobenzene (CDNB) and their failure to bind to immobilized glutathione affinity matrices. Little is known about the number of genes in this class. Recently, Pemble et al. (Biochem J. 300: 271–276, 1994) reported the cDNA cloning of a human Theta-class GST, termed GSTT1. In this study, we describe the molecular cloning of a cDNA encoding a second human Theta-class GST (GSTT2) from a λgt11 human liver 5′-stretch cDNA library. The encoded protein contains 244 amino acids and has 78.3% sequence identity with the rat subunit 12 and only 55.0% identity with human GSTT1. GSTT2 has been mapped to chromosome 22 by somatic cell hybrid analysis. The precise position of the gene was localized to subband 22q11.2 by in situ hybridization. The absence of other regions of hybridization suggests that there are no closely related sequences (e.g., reverse transcribed pseudogenes) scattered throughout the genome and that if there are closely related genes, they must be clustered near GSTT2. Southern blot analysis of human DNA digested with BamHI shows that the size of the GSTT2 gene is relatively small, as the coding sequence falls within a 3.6-kb BamHI fragment.

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Glutathione S-transferases and their implications in the lung diseases asthma and chronic obstructive pulmonary disease: Early life susceptibility?
2021, Redox Biology
Citation Excerpt :
GST Theta (GSTT) is predominantly found in the liver (hepatocytes), and is also expressed in the kidney (renal proximal tubule cells), gastrointestinal tract, and lung [106–108]. The Theta class includes the isoforms GSTT1 and GSTT2, which are both located on chromosome 22 [109,110]. GSTT1 and GSTT2 share 55% amino acid sequence identity.
Our lungs are exposed daily to airborne pollutants, particulate matter, pathogens as well as lung allergens and irritants. Exposure to these substances can lead to inflammatory responses and may induce endogenous oxidant production, which can cause chronic inflammation, tissue damage and remodeling. Notably, the development of asthma and Chronic Obstructive Pulmonary Disease (COPD) is linked to the aforementioned irritants. Some inhaled foreign chemical compounds are rapidly absorbed and processed by phase I and II enzyme systems critical in the detoxification of xenobiotics including the glutathione-conjugating enzymes Glutathione S-transferases (GSTs). GSTs, and in particular genetic variants of GSTs that alter their activities, have been found to be implicated in the susceptibility to and progression of these lung diseases. Beyond their roles in phase II metabolism, evidence suggests that GSTs are also important mediators of normal lung growth. Therefore, the contribution of GSTs to the development of lung diseases in adults may already start in utero, and continues through infancy, childhood, and adult life. GSTs are also known to scavenge oxidants and affect signaling pathways by protein-protein interaction. Moreover, GSTs regulate reversible oxidative post-translational modifications of proteins, known as protein S-glutathionylation. Therefore, GSTs display an array of functions that impact the pathogenesis of asthma and COPD.
In this review we will provide an overview of the specific functions of each class of mammalian cytosolic GSTs. This is followed by a comprehensive analysis of their expression profiles in the lung in healthy subjects, as well as alterations that have been described in (epithelial cells of) asthmatics and COPD patients. Particular emphasis is placed on the emerging evidence of the regulatory properties of GSTs beyond detoxification and their contribution to (un)healthy lungs throughout life. By providing a more thorough understanding, tailored therapeutic strategies can be designed to affect specific functions of particular GSTs.
Genetic polymorphism of glutathione S-transferases: Relevance to neurological disorders
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Chen et al. [58] has studied the frequency of the GSTT1 null genotype in myelodysplastic syndromes and he said 46% of myelodysplastic syndrome patients have null GSTT1 than 16% of controls. The GSTT2 contains 244 amino acids and share 55% similarity in sequence with GSTT1 [59], SNPs in the promoter region of this gene (−537 G→A, −277T→C and −158G→A) may confer greater risk of colorectal cancer [60]. There are two members of omega (ω) class GSTs, they are GSTO1 and GSTO2, those loci exist on chromosome 10 [61], but omega (ω) GST is distinct in case of structure and function from all other GSTs.
Glutathione S-tranferases (GSTs) are phase II drug metabolizing enzymes, they play crucial role in detoxification of environmental pollutants, carcinogens, drugs, xenobiotics and oxidative stress products. Genetic differences in expression and activity of GSTs are due to the existence of polymorphic alleles which encode them. Because of genetic polymorphism the GST activity has altered that lead to the increased susceptibility for toxic chemical compounds. GST genetic polymorphism is the main reason for many neurological dysfunctions. GST has over expressed in epileptic brain and pi (π) GST has used to predict stroke; mu (μ) and pi (π) GST are over expressed in Alzheimer’s disease (AD). Null and single nucleotide polymorphism of GST has associated with many neurodisorders. Over all, it can be concluded that the GST genetic polymorphism has associated with neurodegenerative diseases.
Role of glutathione S-transferases polymorphisms and monocyte CD64 expression in Egyptian patients with systemic lupus erythematosus
2017, Egyptian Rheumatologist
Citation Excerpt :
These enzymes catalyze detoxification of a wide variety of potentially toxic and carcinogenic electrophiles in human environment, by conjugating to glutathione (GSH) [11,12]. Gene clusters of GSTµ (GSTM1, M2, M3, M4, and M5) and GST Θ (GSTT1 and T2) are located on chromosomes 1 and 22, respectively [13]. Independent gene deletions exist at both GSTM1 and GSTT1 loci, resulting in a lack of active protein in 50% and 20% of Caucasians, respectively [14].
To study the genetic variants of glutathione S-transferases and monocyte CD64 expression in systemic lupus erythematosus patients and to evaluate their role in disease susceptibility, activity and damage.
Forty female SLE patients and 40 age matched controls were genotyped for GSTP1, GSTM1 and GSTT1 gene polymorphisms using polymerase chain reaction-restriction fragment length polymorphism, conventional PCR and were assessed for monocyte CD64 expression level using flow cytometry. SLE disease activity index (SLEDAI) and the systemic lupus international. collaborating clinics/damage index (SLICC DI) were considered.
The patients mean age was 28.13 ± 4.56 years and disease duration of 6.4 ± 4.9 with a SLEDAI of 14.4 ± 7.1 and SLICC/DI 3.7 ± 1.5. The frequency of GSTM1 null genotype tended to be higher (55%) in SLE patients compared to the controls (and 42.5%) (p = 0.09). The frequency of GSTT1 null genotype was significantly higher in SLE patients (25%) compared to controls (12.5%) (p < 0.001) and with a 1.7-fold risk. The genotypes frequencies of GSTP1 polymorphism were comparable between SLE patients and controls. The monocyte CD64 expression was significantly increased in the patients (MFI: 46.23 ± 4.56) compared to the control (MFI: 14.05 ± 2.01) (p = 0.001). The GSTM1 and GSTT1 as well as CD64 significantly correlated with the serum creatinine (p = 0.005, p = 0.01 and p-0.001, respectively).
The GST gene polymorphisms together with monocyte CD64 expression level could have a significant relation with SLE and with increased risk in Egyptian patients. The GST gene polymorphisms and monocyte CD64 may form potential biomarkers for renal function.
Molecular cloning and functional characterization of theta class glutathione S-transferase from Apostichopus japonicus
2017, Fish and Shellfish Immunology
Citation Excerpt :
In terms of evolution, biochemistry and catalytic activity, theta class is considered as one of the most interesting GSTs [18]. To date, two types of theta GSTs, GST-θ1 and GST-θ2 have been identified and cloned from human [19,20]. In comparison to other classes, GST-θ shows a specific serine (Ser11) residue in place of the N-terminal tyrosine found in classes of alpha, mu, and phi, which plays a key role in these GSTs for glutathione deprotonation and activation [21,22].
Glutathione S-transferases (GSTs) are the superfamily of multifunctional detoxification isoenzymes and play crucial roles in innate immunity. In the present study, a theta class GST homology was identified from A. japonicus (designated as AjGST-θ) by RACE approaches. The full-length cDNA of AjGST-θ was of 1013 bp encoded a cytosolic protein of 231 amino acids residues. Structural analysis revealed that AjGST-θ processed the characteristic N-terminal GSH-binding site (G-site) and the C-terminal hydrophobic substrate binding site (H-site). Multiple sequence alignment and phylogenetic analysis together supported that AjGST-θ belonged to a new member of theta class GST protein subfamily. Spatial expression analysis revealed that AjGST-θ was ubiquitously expressed in all examined tissues with the larger magnitude in intestine. The Vibrio splendidus challenge in vivo and LPS stimulation in vitro could both significantly up-regulate the mRNA expression of AjGST-θ when compared with control group. The recombinant protein was expressed in Escherichia coli and the purified AjGST-θ showed high activity with GST substrate. Meantime, disc diffusion assay showed that recombinant AjGST-θ protein could markedly improve bacterial growth under Cumene hydroperoxide exposure. More importantly, the recombinant AjGST-θ could effectively prevent primary coelomocytes apoptosis after LPS exposure. Our present findings suggested that AjGST-θ might play significantly roles in the modulation of immune response and protect cells from pathogens infection in A. japonicus.
Glutathione transferases, regulators of cellular metabolism and physiology
2013, Biochimica et Biophysica Acta - General Subjects
Citation Excerpt :
Further investigations are required to fully elucidate the structure/substrate interactions that could underlie the reported associations between GSTP1 genotype, xenobiotic metabolism and cancer susceptibility. Although there are at least 3 Theta class GSTs expressed in the mouse [41], there are only two functional Theta class GST genes in humans (GSTT1 and GSTT2) and both map to chromosome 22q11.2 [125,126]. The locus is however quite complicated because of the frequent deletion of GSTT1 [127] and the inverted duplication of GSTT2 to form a transcribed pseudogene termed GSTT2p or GSTT2b [126].
The cytosolic glutathione transferases (GSTs) comprise a super family of proteins that can be categorized into multiple classes with a mixture of highly specific and overlapping functions.
The review covers the genetics, structure and function of the human cytosolic GSTs with particular attention to their emerging roles in cellular metabolism.
All the catalytically active GSTs contribute to the glutathione conjugation or glutathione dependant-biotransformation of xenobiotics and many catalyze glutathione peroxidase or thiol transferase reactions. GSTs also catalyze glutathione dependent isomerization reactions required for the synthesis of several prostaglandins and steroid hormones and the catabolism of tyrosine. An increasing body of work has implicated several GSTs in the regulation of cell signaling pathways mediated by stress-activated kinases like Jun N-terminal kinase. In addition, some members of the cytosolic GST family have been shown to form ion channels in intracellular membranes and to modulate ryanodine receptor Ca^2 + channels in skeletal and cardiac muscle.
In addition to their well established roles in the conjugation and biotransformation of xenobiotics, GSTs have emerged as significant regulators of pathways determining cell proliferation and survival and as regulators of ryanodine receptors that are essential for muscle function. This article is part of a Special Issue entitled Cellular functions of glutathione.
Genetic polymorphism and function of glutathione S-transferases in tumor drug resistance
2007, Current Opinion in Pharmacology
The human glutathione S-transferase, GSTs, possess both enzymatic and non-enzymatic functions and are involved in many important cellular processes, such as, phase II metabolism, stress response, cell proliferation, apoptosis, oncogenesis, tumor progression and drug resistance. The non-enzymatic functions of GSTs involve their interactions with cellular proteins, such as, JNK, TRAF, ASK, PKC, and TGM2, during which, either the interacting protein partner undergoes functional alteration or the GST protein itself is post-translationally modified and/or functionally altered. The majority of GST genes harbor polymorphisms that influence their transcription and/or function of their encoded proteins. This overview focuses on recent insights into the biology and pharmacogenetics of GSTs as a determinant of cancer drug resistance and response of cancer patients to therapy.

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Molecular cloning of a cDNA and chromosomal localization of a human theta-class glutathione S-transferase gene (GSTT2) to chromosome 22

Abstract

FEBS Lett.

Genomics

Anal. Biochem.

J. Biol. Chem.

FEBS Lett.

Cell

Biochem. Biophys. Res. Commun.

J. Mol. Biol.

J. Mol. Biol.

Biochem. Biophys. Res. Commun.

FEBS Lett.

Genomics

Biochim. Biophys. Acta

Screening lgt recombinant clones by hybridization to single plaques in situ

Science

Genetic heterogeneity of the human glutathione transferases: A complex of gene families

Pharmacol. Ther.

Biochemical genetics of glutathione S-transferase in man

Am. J. Hum. Genet.

Isolation of a cDNA clone and localization of human glutathione S-transferase 2 genes to chromosome band 6p12

Proc. Natl. Acad. Sci. USA