Alteration of metallothionein mRNA in bay scallop Argopecten irradians under cadmium exposure and bacteria challenge.
Metallothionein (MT) is a superfamily of cysteine-rich proteins contributing to metal metabolism, detoxification of heavy metals, and immune response such as protecting against ionizing radiation and antioxidant defense. A metallothionein (designated AiMT2) gene was identified and cloned from bay scallop, Argopecten irradians. The full length cDNA of AiMT2 consisted of an open reading frame (ORF) of 333 bp encoding a protein of 110 amino acids, with nine characteristic Cys-X-Cys, five Cys-X-X-Cys, five Cys-X-X-X-Cys and two Cys-Cys motif arrangements and a conserved structural pattern Cys-x-Cys-x(3)-Cys-Tyr-x(3)-Cys-x-Cys-x(3)-Cys-x-Cys-Arg at the C-terminus. The cloned AiMT showed about 50% identity in the deduced amino acid sequence with previously published MT sequences of mussels and oysters. The conserved structural pattern and the close phylogenetic relationship of AiMT2 shared with MTs from other mollusc especially bivalves indicated that AiMT2 was a new member of molluscan MT family. The mRNA transcripts in hemolymph of AiMT2 under cadmium (Cd) exposure and bacteria challenge were examined by real-time RT-PCR. The mRNA expression of AiMT2 was up-regulated to 3.99-fold at 2 h after Listonella anguillarum challenge, and increased drastically to 66.12-fold and 126.96-fold at 16 and 32 h post-challenge respectively. Cadmium ion exposure could induce the expression of AiMT2, and the expression level increased 2.56-fold and 6.91-fold in hemolymph respectively after a 10-day exposure of 100 microg L(- 1) and 200 microg L(- 1) CdCl(2). The sensitivity of AiMT2 to bacteria challenge and cadmium stress indicated it was a new Cd-dependent MT in bay scallop and also regulated by an immune challenge. The changes in the expression of AiMT2 could be used as an indicator of exposure to metals in pollution monitoring programs and oxidative stress, and bay scallop as a potential sentinel organism for the cadmium contamination in aquatic environment.
Wang
Lingling
L
Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Song
Linsheng
L
Ni
Duojiao
D
Zhang
Huan
H
Liu
Weiqing
W
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000097:blood; D019256:Cadmium Chloride / Q000633:toxicity*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods; D019143:Evolution, Molecular; D006458:Hemolymph / Q000378:metabolism*; D053526:Listonella / Q000472:pathogenicity*; D008668:Metallothionein / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D049897:Pectinidae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D010802:Phylogeny; D012333:RNA, Messenger / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D015415:Biomarkers; D012333:RNA, Messenger; D014874:Water Pollutants, Chemical; D008668:Metallothionein; D019256:Cadmium Chloride