Trichostrongylus vitrinus (Nematoda: Strongylida): molecular characterization and transcriptional analysis of Tv-stp-1, a serine/threonine phosphatase gene.
A full-length cDNA (Tv-stp-1) encoding a serine/threonine protein phosphatase (Tv-STP-1) was isolated from Trichostrongylus vitrinus (order Strongylida), an economically important parasitic nematode of small ruminants. The uninterrupted open reading frame (ORF) of 951 nucleotides encoded a predicted protein of 316 amino acids (aa), containing the characteristic motif [LIVMN]-[KR]-G-N-H-E. Comparison with other sequences in non-redundant databases showed that Tv-STP-1 had significant identities/similarities to those from a range of metazoans and protists. Sequence similarity was most pronounced in the central region of the protein, in which the catalytic activity is inferred to be modulated by eight conserved residues (Asp 61, His 63, Asp 92, Asp 95, Asn 121, His 171, His 246 and Tyr 270), known to coordinate the binding of two metal ions (Mn2+ and Fe2+) in various organisms. Phylogenetic analyses of selected amino acid sequence data using the neighbor-joining and maximum parsimony methods revealed Tv-STP-1 to be most closely related to the glc seven-like phosphatases inferred for genes from the free-living nematode Caenorhabditis elegans and the parasitic nematode Oesophagostomum dentatum (order Strongylida). Comparison of the genomic organization of the full-length Tv-stp-1 gene with related molecules from other nematodes revealed substantial variation in the lengths and numbers of the exons and introns. The entire genes Tv-stp-1 (5041-5362 bp; 10 exons and 9 introns) and Od-mpp-1 (10,271 bp; 8 exons and 9 introns) from the parasitic nematodes T. vitrinus and O. dentatum were considerably longer than the C. elegans genes (1222-1603 bp; 3-7 exons and 2-6 introns). Transcriptional analysis by reverse transcription polymerase chain reaction (RT-PCR) showed that Tv-stp-1 was transcribed in adult males of T. vitrinus, but not in the adult female or in any larval stages of this species. In spite of considerable variation at the genomic level, the findings of the present study suggest that there is relative conservation in features and function of the serine/threonine protein phosphatase characterized among T. vitrinus, O. dentatum and C. elegans, which should have implications for exploring molecular reproductive and developmental processes in strongylid nematodes of socio-economic importance.
Hu
Min
M
Department of Veterinary Science, The University of Melbourne, 250 Princes Highway, Werribee, Victoria 3030, Australia.
Abs EL-Osta
Youssef G
YG
Campbell
Bronwyn E
BE
Boag
Peter R
PR
Nisbet
Alasdair J
AJ
Beveridge
Ian
I
Gasser
Robin B
RB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D018509:DNA, Helminth / Q000737:chemistry* / Q000302:isolation & purification; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000235:genetics*; D010802:Phylogeny; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology; D014158:Transcription, Genetic; D014253:Trichostrongylosis / Q000469:parasitology / Q000662:veterinary; D014254:Trichostrongylus / Q000145:classification / Q000201:enzymology / Q000235:genetics*
D018076:DNA, Complementary; D018509:DNA, Helminth; D018510:RNA, Helminth; D010749:Phosphoprotein Phosphatases