Tolliod-like gene in Crassostrea ariakensis: Molecular cloning, structural characterization and expression by RLO stimulation.
The BMP1/TLD-like proteinases are pleiotropic, astacin-like metalloproteinases. They play central roles in regulating the formation of the extracellular matrix (ECM) and signaling through various TGFbeta-like proteins in morphogenetic and homeostatic events. Here we describe the cloning, structural characterization and expression of Tolloid-like gene in the oyster, Crassostrea ariakensis (CaTLL). The full-length cDNA of CaTLL spans 3492 nucleotides including an open reading frame of 2811 nucleotides which encodes a hypothetical protein of 936 amino acids, with a molecular mass of approximately 103 kDa. The CaTLL molecule possessed structural features of several motifs including an N-terminal signal peptide sequence, a prodomain with an RTRR motif, an astacin-like domain that contains a conserved zinc-binding motif HELGHVIGFWHEH, five CUBs and two EGF domains with the arrangement CUB-CUB-EGF-CUB-EGF-CUB-CUB. The proteolytic domain of Ca-Tolloid shares more than 30% identity with other astacins of various animals from squail to mammals, indicating its conserved catalytic ability. RT-PCR and quantitative real-time PCR analyses revealed that CaTLL showed the lowest expression level in hemocytes of normal groups, but was affected significantly by the challenge of an obligate intracellular Gram-negative bacterium, Rickettsia-like organisms, suggesting that Ca-Tolloid might be involved in the molluscan immune response, and its function is more diverse than previously assumed.
Yang
Shoubao
S
Laboratory of Marine Life Science and Technology, College of Animal Sciences, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang 310029, China; Shaoxing College of Arts and Sciences, Zhejiang, China.
Wu
Xinzhong
X
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D049896:Crassostrea / Q000201:enzymology* / Q000235:genetics*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D055394:Tolloid-Like Metalloproteinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D055394:Tolloid-Like Metalloproteinases