Identification and expression analysis of the Steinernema carpocapsae elastase-like serine protease gene during the parasitic stage.
A cDNA encoding elastase was isolated from Steinernema carpocapsae by suppression subtractive hybridization and rapid amplification of 5' cDNA ends. The predicted protein contained a 19-aa signal peptide, a 44-aa N-terminal propeptide, and a 264-aa mature protein with a predicted molecular mass of 28,949 Da and a theoretical pI of 8.88. BLAST analysis showed 27-35% amino acid sequence identity to serine proteases from insects, mammals, fish and other organisms. The Sc-ela gene contains three exons and two introns with at least two copies in the S. carpocapsae genome. Expression analysis indicated that the Sc-ela gene was upregulated during the initial parasitic stage. Sequence comparison and evolutionary marker analysis revealed that Sc-ELA was a member of the elastase serine protease family with potential degradative, developmental and fibrinolytic activities. Homology modeling showed that Sc-ELA adopts a two beta-barrel fold typical of trypsin-like serine proteases, and phylogenetic analysis indicates that Sc-ELA branched off early during elastase evolution.
Hao
You-Jin
YJ
CIRN and Department of Biology, University of Azores, 9501-801 Ponda Delgada, Azores, Portugal. haoyoujin@hotmail.com
Montiel
Rafael
R
Nascimento
Gisela
G
Toubarro
Duarte
D
Simoes
Nelson
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D006790:Host-Parasite Interactions; D008969:Molecular Sequence Data; D010572:Pest Control, Biological; D010802:Phylogeny; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017168:Rhabditida / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment; D057057:Serine Proteases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D013379:Substrate Specificity
D018076:DNA, Complementary; D018510:RNA, Helminth; D057057:Serine Proteases