Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca<sup>2+</sup> out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca<sup>...Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca<sup>2+</sup> out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca<sup>2+</sup>-ATPase gene was isolated from the gill of <em>Hyriopsis cumingii</em> (HcPMCA) by using SMART RACE technique. The entire cDNA was 5230 bp, including a 417-bp 5'-UTR, a 3588-bp ORF and a 1225-bp 3'-UTR, encoding a 1195-amino acid protein, and no putative signal peptide was predicted. Compared with PMCA homologs from seawater mollusks, HcPMCA had high similarity with them in both sequence and structure. Tissue-specific expression analysis revealed that HcPMCA mRNA was detected in all the sampled tissues, but was prominently expressed in the gill and mantle. When exposed to a serie of increasing Ca<sup>2+</sup> that lasted for 7 days, the mRNA expression of HcPMCA in the mantle was slightly downregulated, but peaked at 60 mg/L. Moreover, the temporal expression of HcPMCA transcripts in the mantle after 60 mg/L Ca<sup>2+</sup> exposure was shown to be bell-shaped, which was slightly downregulated at 24 h, but upregulated from 24 h to 48 h post-treatment, peaking at 48 h. The result of present study provides useful information for further studies on function and regulation mechanism of HcPMCA gene.展开更多
In triangle sail mussel(Hyriopsis cumingii),shell biomineralization is a complicated process that involves multiple gene products.Shell matrix proteins are involved in the formation of the organic framework and play a...In triangle sail mussel(Hyriopsis cumingii),shell biomineralization is a complicated process that involves multiple gene products.Shell matrix proteins are involved in the formation of the organic framework and play an important role in the regulation of calcium carbonate deposition.In this study,A new shell matrix protein gene Hc-transgelin was identified in H.cumingii.The full-length cDNA of Hc-transgelin was 1200 bp,including a 501 bp open reading frame,which encoded 166 amino acids.Hc-transgelin is rich in lysine,it accounts for 11.40%of the protein.The predicted transgelin protein contained a conserved calmodulin homologous domain.A tissue-specific expression assay indicated that Hc-transgelin exhibited significantly highest expression in the mantle.Furthermore,Hc-transgelin in situ hybridization detected positive signals at the edge of the mantle outer fold,where nacre and prismatic layer biomineralization occur.An RNA interference assay showed that the shape of aragonite flakes in nacre changed and their growth was inhibited,and cracks appeared in the prismatic layer organic sheath when the expression of Hc-transgelin was suppressed.In a shell repair assay,a higher expression of Hc-transgelin appeared from day 12 to day 25 when the nacre accumulated quickly.These findings indicate that Hc-transgelin may be involved in the formation of aragonite flakes in the nacre and play a role in the formation of the organic sheath in the prismatic layer of the shell.This study provides new insights into the role of the Hc-transgelin gene and also contributes to the molecular understanding of mollusk shell formation.展开更多
文摘Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca<sup>2+</sup> out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca<sup>2+</sup>-ATPase gene was isolated from the gill of <em>Hyriopsis cumingii</em> (HcPMCA) by using SMART RACE technique. The entire cDNA was 5230 bp, including a 417-bp 5'-UTR, a 3588-bp ORF and a 1225-bp 3'-UTR, encoding a 1195-amino acid protein, and no putative signal peptide was predicted. Compared with PMCA homologs from seawater mollusks, HcPMCA had high similarity with them in both sequence and structure. Tissue-specific expression analysis revealed that HcPMCA mRNA was detected in all the sampled tissues, but was prominently expressed in the gill and mantle. When exposed to a serie of increasing Ca<sup>2+</sup> that lasted for 7 days, the mRNA expression of HcPMCA in the mantle was slightly downregulated, but peaked at 60 mg/L. Moreover, the temporal expression of HcPMCA transcripts in the mantle after 60 mg/L Ca<sup>2+</sup> exposure was shown to be bell-shaped, which was slightly downregulated at 24 h, but upregulated from 24 h to 48 h post-treatment, peaking at 48 h. The result of present study provides useful information for further studies on function and regulation mechanism of HcPMCA gene.
基金supported by the National Key R&D Program of China (2018YFD0901406)the National Natural Science Foundation of China (31872565)+1 种基金the earmarked fund for CARS (CARS-49)the Sponsored by Program of Shanghai Academic Research Leader (19XD1421500).
文摘In triangle sail mussel(Hyriopsis cumingii),shell biomineralization is a complicated process that involves multiple gene products.Shell matrix proteins are involved in the formation of the organic framework and play an important role in the regulation of calcium carbonate deposition.In this study,A new shell matrix protein gene Hc-transgelin was identified in H.cumingii.The full-length cDNA of Hc-transgelin was 1200 bp,including a 501 bp open reading frame,which encoded 166 amino acids.Hc-transgelin is rich in lysine,it accounts for 11.40%of the protein.The predicted transgelin protein contained a conserved calmodulin homologous domain.A tissue-specific expression assay indicated that Hc-transgelin exhibited significantly highest expression in the mantle.Furthermore,Hc-transgelin in situ hybridization detected positive signals at the edge of the mantle outer fold,where nacre and prismatic layer biomineralization occur.An RNA interference assay showed that the shape of aragonite flakes in nacre changed and their growth was inhibited,and cracks appeared in the prismatic layer organic sheath when the expression of Hc-transgelin was suppressed.In a shell repair assay,a higher expression of Hc-transgelin appeared from day 12 to day 25 when the nacre accumulated quickly.These findings indicate that Hc-transgelin may be involved in the formation of aragonite flakes in the nacre and play a role in the formation of the organic sheath in the prismatic layer of the shell.This study provides new insights into the role of the Hc-transgelin gene and also contributes to the molecular understanding of mollusk shell formation.
