Molecular characterisation of a male-specific serine/threonine phosphatase from Oesophagostomum dentatum (Nematoda: Strongylida), and functional analysis of homologues in Caenorhabditis elegans.
A male-specifically expressed sequence tag was used as a probe to screen adult male Oesophagostomum dentatum (Nematoda; Strongylida) gene libraries. The cDNA clones isolated coded for a serine/threonine protein phosphatase with approximately 85% identity to two Caenorhabditis elegans proteins implicated in reproduction. However, the genomic structures for the two species were distinct, in that the O. dentatum gene contained seven introns, whereas the C. elegans homologues contained three (two of which were conserved between the two nematodes). The promoters of all three nematode genes contained two putative GATA motifs separated by six to seven nucleotides and located within 100 nucleotides of the predicted transcriptional start site. RNA interference (RNAi) experiments in C. elegans, targetting the two homologues, revealed a consistent reduction in the number of progeny produced by treated worms, indicating a functional role in reproduction. Expression of green fluorescent protein, directed by the putative promoters for the C. elegans phosphatase genes, was analysed in transgenic C. elegans. The present results suggest that there is a significant degree of conservation between O. dentatum and C. elegans in the features and function of the serine/threonine protein phosphatase characterised, which should have implications for detailed investigations into molecular reproductive processes of some parasitic nematodes.
Boag
Peter R
PR
Victorian Institute of Animal Science, Victoria 3049, Attwood, Australia.
Ren
Peifeng
P
Newton
Susan E
SE
Gasser
Robin B
RB
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000235:genetics*; D020869:Gene Expression Profiling; D017238:Genes, Helminth; D049452:Green Fluorescent Proteins; D007438:Introns; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009815:Oesophagostomum / Q000201:enzymology / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010749:Phosphoprotein Phosphatases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012098:Reproduction; D016415:Sequence Alignment
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D010749:Phosphoprotein Phosphatases