Identification of stress-responsive genes in Caenorhabditis elegans using RT-PCR differential display.
In order to identify genes that are differentially expressed as a consequence of oxidative stress due to paraquat we used the differential display technique to compare mRNA expression patterns in Caenorhabditis elegans . A C.elegans mixed stage worm population and a homogeneous larval population were treated with 100 mM paraquat, in parallel with controls. Induction of four cDNA fragments, designated L-1, M-47, M-96 and M-132, was confirmed by Northern blot analysis with RNA from stressed and unstressed worm populations. A 40-fold increase in the steady-state mRNA level in the larval population was observed for the L-1/M-47 gene, which encodes the detoxification enzyme glutathione S-transferase. A potential stress-responsive transcription factor (M-132) with C2H2-type zinc finger motifs and an N-terminal leucine zipper domain was identified. The M-96 gene encodes a novel stress-responsive protein. Since paraquat is known to generate superoxide radicals in vivo , the response of the C.elegans superoxide dismutase (SOD) genes to paraquat was also investigated in this study. The steady-state mRNA levels of the manganese-type and the copper/zinc-type SODs increased 2-fold in the larval population in response to paraquat, whereas mixed stage populations did not show any apparent increase in the levels of these SOD mRNAs.
Tawe
W N
WN
Department of Biochemistry, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht Strasse 74, 20359 Hamburg, Germany.
Eschbach
M L
ML
Walter
R D
RD
Henkle-Dührsen
K
K
D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D018076:DNA, Complementary; D005786:Gene Expression Regulation / Q000187:drug effects*; D017238:Genes, Helminth; D005982:Glutathione Transferase / Q000096:biosynthesis; D007814:Larva; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D010269:Paraquat / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods; D018510:RNA, Helminth / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D013312:Stress, Physiological; D013482:Superoxide Dismutase / Q000096:biosynthesis; D014157:Transcription Factors / Q000096:biosynthesis* / Q000737:chemistry; D014158:Transcription, Genetic; D016335:Zinc Fingers
D018076:DNA, Complementary; D018510:RNA, Helminth; D012333:RNA, Messenger; D014157:Transcription Factors; D013482:Superoxide Dismutase; D005982:Glutathione Transferase; D010269:Paraquat