Abstract
The blood clam Tegillarca granosa, a eukaryotic bottom-dwelling bivalve species has a strong ability to tolerate and accumulate cadmium. In our previous study, Nfu1 (iron-sulfur cluster scaffold protein), which is involved in Fe-S cluster biogenesis, was shown to be significantly up-regulated under Cd stress, as determined by proteomic analysis. To investigate the function of Nfu1 in cadmium (Cd) detoxification, the function of blood clam Nfu1 (designated as Tg-Nfu1) was investigated by integrated molecular and protein approaches. The full-length cDNA of Tg-Nfu1 is 1167 bp and encodes a protein of 272 amino acid residues. The deduced Tg-Nfu1 protein is 30 kDa contains a conserved Nfu-N domain and a Fe-S cluster binding motif (C-X-X-C). qRT-PCR analysis revealed that Tg-Nfu1 was ubiquitously expressed in all examined tissues; it was up-regulated in the hepatopancreas and gill, and kept a high level from 9 to 24 h after Cd exposure (250 μg/L). Western blot analysis further revealed that the Tg-Nfu1 protein was also highly expressed in the hepatopancreas and gill after 24 h of Cd stress. Further functional analysis showed that the production of ROS was increased and Cu/ZnSOD activity was inhibited in blood clam, treated with the specific Nfu1 siRNA and Cd stress, respectively. These results suggest that Tg-Nfu1 could protect blood clam from oxidative damage caused by Cd stress.
Highlights
In recent years, with the development of coastal economies, pollution of marine ecosystems by heavy metals has caused damage as a result of industrial activity but has seriously limited the sustainable development of aquaculture, on the east coast of China (Wu et al, 2012; Wang et al, 2013)
We found that Nfu1 protein expression was significantly increased in blood clam under Cd stress, as measured by iTRAQ proteome analysis (Bao et al, 2016)
The 3′- and 5′- ends of Nfu1 were cloned using the SMARTRACE approach
Summary
With the development of coastal economies, pollution of marine ecosystems by heavy metals has caused damage as a result of industrial activity but has seriously limited the sustainable development of aquaculture, on the east coast of China (Wu et al, 2012; Wang et al, 2013). The pollution of Cadmium (Cd) is increasing serious in the East China Sea due to the drainage of industrial sewage (Bao et al, 2014). As a benthic filter feeder, blood clam has higher Cd accumulation capacity and tolerance to the metal’s toxicity than oysters and mussels (Gong et al, 2011), and it is a great object for studying resistance to various natural stresses, such as hypoxia, heavy metals and pollutants (Bao et al, 2014). Numerous studies have reported the toxicological mechanism or enrichment of Cd binding proteins, for example, metallothionein (MT)
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