Alternative mRNAs arising from trans-splicing code for mitochondrial and cytosolic variants of Echinococcus granulosus thioredoxin Glutathione reductase.
Thioredoxin and glutathione systems are the major thiol-dependent redox systems in animal cells. They transfer via the reversible oxidoreduction of thiols the reducing equivalents of NADPH to numerous substrates and substrate reductases and constitute major defenses against oxidative stress. In this study, we cloned from the helminth parasite Echinococcus granulosus two trans-spliced mRNA variants that encode thioredoxin glutathione reductases (TGR). These variants code for mitochondrial and cytosolic selenocysteine-containing isoforms that possess identical glutaredoxin (Grx) and thioredoxin reductase (TR) domains and differ exclusively in their N termini. Western blot analysis of subcellular fractions with specific anti-TGR antibodies showed that TGR is present in both compartments. The biochemical characterization of the native purified TGR suggests that the Grx and TR domains of the enzyme can function either coupled or independently of each other, because the Grx domain can accept electrons from either TR domains or the glutathione system and the TR domains can transfer electrons to either the fused Grx domain or to E. granulosus thioredoxin.
Agorio
Astrid
A
Cátedra de Inmunología, Facultad de Química/Ciencias, Universidad de la República, Avenida Alfredo Navarro 3051, Piso 2, CP 11.600, Montevideo, Uruguay.
Chalar
Cora
C
Cardozo
Soledad
S
Salinas
Gustavo
G
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003600:Cytosol / Q000201:enzymology; D017931:DNA Primers; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004446:Echinococcus / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D005091:Exons; D014644:Genetic Variation; D006801:Humans; D007700:Kinetics; D051379:Mice; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D018510:RNA, Helminth / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D051381:Rats; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D009097:Multienzyme Complexes; D018510:RNA, Helminth; D012333:RNA, Messenger; D009247:NADH, NADPH Oxidoreductases; C466433:thioredoxin glutathione reductase