Decreased mitochondrial hydrogen peroxide release in transgenic Drosophila melanogaster expressing intramitochondrial catalase

被引:19
作者
Kwong, LK [1 ]
Mockett, RJ [1 ]
Bayne, ACV [1 ]
Orr, WC [1 ]
Sohal, RS [1 ]
机构
[1] So Methodist Univ, Dept Biol Sci, Dallas, TX 75275 USA
关键词
catalase; mitochondria; transgenic; chimeric proteins; ectopic expression; signal peptides; oxidative stress; H2O2;
D O I
10.1006/abbi.2000.2093
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The objective of this study was to develop strategies for manipulating oxidative stress transgenically in a multicellular organism. Ectopic catalase was introduced into the mitochondrial matrix, which is the main intracellular site of H2O2 formation and where catalase is normally absent. Transgenic Drosophila melanogaster were generated by microinjection of a P element construct, containing the genomic catalase sequence of Drosophila, with the mitochondrial leader sequence of ornithine aminotransferase inserted upstream of the coding region, Total catalase activities in whole-body homogenates of 10-day-old flies from four transgenic lines were similar to 30-160% higher than those from the parental and four vector-only control lines. Expression of catalase in the mitochondrial matrix was confirmed by immunoblotting and catalase activity assays. Mitochondrial release of H2O2 was decreased by similar to 90% in the transgenic lines when compared to levels in vector-only controls. This in vivo system provides a novel model for examining the functional significance of decreased mitochondrial H2O2 release. (C) 2000 Academic Press.
引用
收藏
页码:303 / 308
页数:6
相关论文
共 38 条
[1]   Overexpression of catalase in cytosolic or mitochondrial compartment protects HepG2 cells against oxidative injury [J].
Bai, JX ;
Rodriguez, AM ;
Melendez, JA ;
Cederbaum, AI .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (37) :26217-26224
[2]   ESCHERICHIA-COLI IRON SUPEROXIDE-DISMUTASE TARGETED TO THE MITOCHONDRIA OF YEAST-CELLS PROTECTS THE CELLS AGAINST OXIDATIVE STRESS [J].
BALZAN, R ;
BANNISTER, WH ;
HUNTER, GJ ;
BANNISTER, JV .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (10) :4219-4223
[3]   MITOCHONDRIAL GENERATION OF HYDROGEN-PEROXIDE - GENERAL PROPERTIES AND EFFECT OF HYPERBARIC-OXYGEN [J].
BOVERIS, A ;
CHANCE, B .
BIOCHEMICAL JOURNAL, 1973, 134 (03) :707-716
[4]   INTRONS INCREASE TRANSCRIPTIONAL EFFICIENCY IN TRANSGENIC MICE [J].
BRINSTER, RL ;
ALLEN, JM ;
BEHRINGER, RR ;
GELINAS, RE ;
PALMITER, RD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (03) :836-840
[5]   HYDROPEROXIDE METABOLISM IN MAMMALIAN ORGANS [J].
CHANCE, B ;
SIES, H ;
BOVERIS, A .
PHYSIOLOGICAL REVIEWS, 1979, 59 (03) :527-605
[6]   Introns boost transgene expression in Drosophila melanogaster [J].
Duncker, BP ;
Davies, PL ;
Walker, VK .
MOLECULAR & GENERAL GENETICS, 1997, 254 (03) :291-296
[7]   Microinjection of catalase cDNA prevents hydrogen peroxide-induced motoneuron death [J].
Herpers, BL ;
Schrama, LH ;
Kaal, ECA ;
Joosten, EAJ ;
Bär, PRD .
NEUROREPORT, 1999, 10 (12) :2647-2650
[9]   THE 9 AMINO-TERMINAL RESIDUES OF DELTA-AMINOLEVULINATE SYNTHASE DIRECT BETA-GALACTOSIDASE INTO THE MITOCHONDRIAL MATRIX [J].
KENG, T ;
ALANI, E ;
GUARENTE, L .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (02) :355-364
[10]  
KOBAYASHI T, 1995, AM J HUM GENET, V57, P284