Cloning and characterisation of a Heterodera glycines aminopeptidase cDNA.
An aminopeptidase full-length cDNA (Hg-amp-1) was cloned from the adult female soybean cyst nematode Heterodera glycines by heterologous screening of a cDNA library with a Caenorhabditis elegans EST sequence. The predicted open reading frame encoded an 882-amino acid protein containing the conserved zinc-binding domain and GAMEN motif that are characteristic of M1 family aminopeptidases. The putative protein lacks any subcellular targeting signals and displays strong similarity to puromycin-sensitive aminopeptidases from C. elegans, Drosophila and mammals. Hg-amp-1 is expressed in juvenile nematodes and both male and female adults, with highest expression in gravid females. In situ mRNA hybridisation localised the Hg-amp-1 transcript to the genital primordium of pre-parasitic juvenile nematodes and the reproductive tract of adult females. Suppression of Hg-amp-1 transcript level by RNA-interference led to a 61% reduction in the number of female nematodes parasitising soybean roots 21 days post infection with infective juvenile nematodes that had been exposed to double-stranded RNA.
Lilley
Catherine J
CJ
Centre for Plant Sciences, University of Leeds, LS2 9JT, UK. c.j.lilley@leeds.ac.uk
Goodchild
Sarah A
SA
Atkinson
Howard J
HJ
Urwin
Peter E
PE
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D017173:Caenorhabditis elegans / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D018509:DNA, Helminth / Q000235:genetics*; D004330:Drosophila / Q000235:genetics; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D017403:In Situ Hybridization; D008018:Life Cycle Stages; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009349:Nematode Infections / Q000201:enzymology; D010935:Plant Diseases / Q000469:parasitology; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000469:parasitology; D014431:Tylenchoidea / Q000201:enzymology* / Q000235:genetics
D018076:DNA, Complementary; D018509:DNA, Helminth; D000626:Aminopeptidases