title: Two isoforms of Saccharomyces cerevisiae glutaredoxin 2 are expressed in vivo and localize to different subcellular compartments.
abstract: Glutaredoxin (Grx)2 from Saccharomyces cerevisiae is a member of the two-cysteine (dithiol) subfamily of Grxs involved in the defence against oxidative stress in yeast. Recombinant yeast Grx2p, expressed in Escherichia coli, behaves as a 'classical' Grx that efficiently catalyses the reduction of hydroxyethyl disulphide by GSH. Grx2p also catalyses the reduction of GSSG by dihydrolipoamide with even higher efficiency. Western blot analysis of S. cerevisiae crude extracts identifies two isoforms of Grx2p of 15.9 and 11.9 kDa respectively. The levels of these two isoforms reach a peak during the exponential phase of growth in normal yeast extract/peptone/dextrose ('YPD') medium, with the long form predominating over the short one. From immunochemical analysis of subcellular fractions, it is shown that both isoforms are present in mitochondria, but only the short one is detected in the cytosolic fraction. On the other hand, only the long form is prominent in microsomes. Mitochondrial isoforms should represent the processed and unprocessed products of an open reading frame (YDR513W), with a putative start codon 99 bp upstream of the GRX2 start codon described thus far. These results indicate that GRX2 contains two in-frame start codons, and that translation from the first AUG results in a product that is targeted to mitochondria. The cytosolic form would result either by initiation from the second AUG, or by differential processing of one single translation product.
authors.affiliation: Departamento de Biología Experimental, Facultad de Ciencias Experimentales, University of Jaén, 23071-Jaén, Spain.
authors.forename: José R
authors.initials: JR
authors.lastname: Pedrajas
authors.forename: Pablo
authors.initials: P
authors.lastname: Porras
authors.forename: Emilia
authors.initials: E
authors.lastname: Martínez-Galisteo
authors.forename: C Alicia
authors.initials: CA
authors.lastname: Padilla
authors.forename: Antonio
authors.initials: A
authors.lastname: Miranda-Vizuete
authors.forename: J Antonio
authors.initials: JA
authors.lastname: Bárcena
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics; D054477:Glutaredoxins; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010088:Oxidoreductases; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: C516005:GLRX protein, human; C516008:GLRX2 protein, human; C516007:Glrx protein, rat; C516009:Glrx2 protein, mouse; C516010:Glrx2 protein, rat; D054477:Glutaredoxins; D007527:Isoenzymes; D011506:Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases