title: Pepsin-like aspartic protease (Sc-ASP155) cloning, molecular characterization and gene expression analysis in developmental stages of nematode Steinernema carpocapsae.
abstract: Steinernema carpocapsae is an insect parasitic nematode associated with the bacterium Xenorhabdus nematophila. These symbiotic complexes are virulent against the insect host. Many protease genes were shown previously to be induced during parasitism, including one predicted to encode an aspartic protease, which was cloned and analyzed in this study. A cDNA encoding Sc-ASP155 was cloned based on the EST fragment. The full-length cDNA of Sc-ASP155 consists of 955 nucleotides with multiple domains, including a signal peptide (aa1-15), a pro-peptide region (aa16-45), and a typical catalytic aspartic domain (aa71-230). The putative 230 amino acid residues have a calculated molecular mass of 23,812Da and a theoretical pI of 5.01. Sc-ASP155 blastp analysis showed 40-62% amino acid sequence identity to aspartic proteases from parasitic and free-living nematodes. Expression analysis showed that the sc-asp155 gene was up-regulated during the initial parasitic stage, especially in L3 gut and 6h induced nematodes. Sequence comparison revealed that Sc-ASP155 was a member of an aspartic protease family and phylogenetic analysis indicated that Sc-ASP155 was clustered with Sc-ASP113. In situ hybridization showed that sc-asp155 was expressed in subventral cells. Additionally, we determined that sc-asp155 is a single-copy gene in S. carpocapsae. Homology modeling showed that Sc-ASP155 adopts a typical aspartic protease structure. The up-regulated Sc-ASP155 expression revealed that this protease could play a role in the parasitic process. In this study, we have cloned the gene and determined the expression of the pepsin-like aspartic protease Sc-ASP155 in S. carpocapsae.
authors.affiliation: CIRN and Department of Biology, University of Azores, 9501-855 Ponta Delgada, Azores, Portugal. natesanbala@uac.pt
authors.forename: Natesan
authors.initials: N
authors.lastname: Balasubramanian
authors.forename: Gisela
authors.initials: G
authors.lastname: Nascimento
authors.forename: Ricardo
authors.initials: R
authors.lastname: Ferreira
authors.forename: Monica
authors.initials: M
authors.lastname: Martinez
authors.forename: Nelson
authors.initials: N
authors.lastname: Simões
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057055:Aspartic Acid Proteases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006790:Host-Parasite Interactions; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009036:Moths / Q000469:parasitology*; D010434:Pepsin A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D018510:RNA, Helminth / Q000235:genetics; D017168:Rhabditida / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000382:microbiology; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013559:Symbiosis; D020593:Xenorhabdus / Q000502:physiology
chemical_list: D015801:Helminth Proteins; D018510:RNA, Helminth; D057055:Aspartic Acid Proteases; D010434:Pepsin A