Molecular evidence for a Fe-hydrogenase in the green alga Scenedesmus obliquus.
Fe-hydrogenase-specific degenerate primers were used in polymerase chain reactions with single-stranded Scenedesmus obliquus cDNA as template. A resulting 435-bp fragment was used to screen a cDNA library from S. obliquus. A 1.6-kb cDNA clone, containing the 3'-end of an open reading frame, was obtained. Its deduced amino acid sequence contains the conserved H-cluster motives unique for Fe-hydrogenases. Any other FeS-clusters seem to be absent. Phylogenetically, the enzyme is closely related to the Trichomonas vaginalis Fe-hydrogenase. Northern blot analysis shows that the Fe-hydrogenase is constitutively expressed in S. obliquus. Southern blot analysis of plastidic, mitochondrial, and nuclear DNA reveals that the enzyme is a single-copy gene encoded in the nucleus. Effects of transcriptional and translational inhibitors on H2-metabolism reveal the involvement of a chloroplast-encoded protein.
Wünschiers
R
R
Fachbereich Biologie/Botanik, Philipps-Universität, Karl-von-Frisch-Str., 35032 Marburg, Germany. wuenschi@yahoo.com
Stangier
K
K
Senger
H
H
Schulz
R
R
D000595:Amino Acid Sequence; D000693:Anaerobiosis; D001483:Base Sequence; D000460:Chlorophyta / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics; D018628:Gene Dosage; D015723:Gene Library; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000235:genetics* / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D014176:Protein Biosynthesis; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D015342:DNA Probes; D018076:DNA, Complementary; D007506:Iron-Sulfur Proteins; D006859:Hydrogen; C118830:iron hydrogenase; D006864:Hydrogenase