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Endoplasmic Reticulum Stress-Related Genes in Yellow Catfish Pelteobagrus fulvidraco: Molecular Characterization, Tissue Expression, and Expression Responses to Dietary Copper Deficiency and Excess.

Song YF, Luo Z, Huang C, Chen QL, Pan YX, Xu YH - G3 (Bethesda) (2015)

Bottom Line: The predicted amino acid sequences for the yellow catfish grp78, crt, perk, ire-1α, and atf-6α revealed that the proteins contained all of the structural features that were characteristic of the five genes in other species, including the KDEL motif, signal peptide, sensor domain, and effector domain. mRNAs of the five genes mentioned above were expressed in various tissues, but their mRNA levels varied among tissues.For the first time, our study cloned grp78, crt, perk, ire-1α, and atf-6α genes in yellow catfish and demonstrated their differential expression among tissues.Moreover, the present study also indicated differential regulation of these ER stress-related genes by dietary Cu deficiency and excess, which will be beneficial for us to evaluate effects of dietary Cu levels in fish at the molecular level, based on the upstream pathway of lipid metabolism (the ER) and thus provide novel insights regarding the nutrition of Cu in fish.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Fishery College, Huazhong Agricultural University, Wuhan 430070, China Freshwater Aquaculture Collaborative Innovative Center of Hubei Province, Wuhan 430070, China.

No MeSH data available.


(A) ClustalX alignment of the deduced amino acid sequence of crt from P. fulvidraco and other species. The identical residues are shaded dark gray. Characteristic features are denoted as follows: SP (signal peptide) brace; N-glycosylation site, triangle; C-terminal KDEL motif, an ER-retrieval sequence, asterisk; N domain, dashed line; P domain, solid line; C domain, dotted line; triplicate repeats A, solid lines box; triplicate repeats B, dotted lines box. Deduced amino acid sequences were obtained from D. rerio (GenBank accession no. NM_131047) and H. sapiens (NM_004343). (B) The predicted peptide features of crt. SP, signal peptide.
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fig2: (A) ClustalX alignment of the deduced amino acid sequence of crt from P. fulvidraco and other species. The identical residues are shaded dark gray. Characteristic features are denoted as follows: SP (signal peptide) brace; N-glycosylation site, triangle; C-terminal KDEL motif, an ER-retrieval sequence, asterisk; N domain, dashed line; P domain, solid line; C domain, dotted line; triplicate repeats A, solid lines box; triplicate repeats B, dotted lines box. Deduced amino acid sequences were obtained from D. rerio (GenBank accession no. NM_131047) and H. sapiens (NM_004343). (B) The predicted peptide features of crt. SP, signal peptide.

Mentions: In the present study, by RT-PCR, 3′- and 5′-RACE PCR, we obtained the full-length cDNA sequences of grp78, crt, perk, ire-1α, and atf-6α genes, which were shown to be 2544 bp, 1678 bp, 3776 bp, 3440 bp, and 2785 bp, respectively (Table 4). Sequencing revealed that their cDNAs included an ORF of 1956 bp, 1254 bp, 3240 bp, 3113 bp, and 1971 bp, respectively, encoding a protein of 652 amino acids, 418 amino acids, 1080 amino acids, 1038 amino acids, and 657 amino acids, respectively. The protein sequence of P. fulvidraco grp78 possessed all the characteristic features of grp78, including signal peptide, C-terminal KDEL motif, ATPase domain, and substrate-binding domain (Figure 1). The protein sequence of yellow catfish crt possessed all the characteristic features of crt, including signal peptide, N-glycosylation site, and C-terminal KDEL motif (Figure 2). The predicted amino acid sequence for the yellow catfish perk, ire-1α, and atf-6α possessed all the characteristic features of ER stress–sensing genes, including signal peptide, transmembrane domain, protein kinase, ribonuclease, sensor domain, and effector domain (Figure 3, Figure 4, and Figure 5).


Endoplasmic Reticulum Stress-Related Genes in Yellow Catfish Pelteobagrus fulvidraco: Molecular Characterization, Tissue Expression, and Expression Responses to Dietary Copper Deficiency and Excess.

Song YF, Luo Z, Huang C, Chen QL, Pan YX, Xu YH - G3 (Bethesda) (2015)

(A) ClustalX alignment of the deduced amino acid sequence of crt from P. fulvidraco and other species. The identical residues are shaded dark gray. Characteristic features are denoted as follows: SP (signal peptide) brace; N-glycosylation site, triangle; C-terminal KDEL motif, an ER-retrieval sequence, asterisk; N domain, dashed line; P domain, solid line; C domain, dotted line; triplicate repeats A, solid lines box; triplicate repeats B, dotted lines box. Deduced amino acid sequences were obtained from D. rerio (GenBank accession no. NM_131047) and H. sapiens (NM_004343). (B) The predicted peptide features of crt. SP, signal peptide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4592992&req=5

fig2: (A) ClustalX alignment of the deduced amino acid sequence of crt from P. fulvidraco and other species. The identical residues are shaded dark gray. Characteristic features are denoted as follows: SP (signal peptide) brace; N-glycosylation site, triangle; C-terminal KDEL motif, an ER-retrieval sequence, asterisk; N domain, dashed line; P domain, solid line; C domain, dotted line; triplicate repeats A, solid lines box; triplicate repeats B, dotted lines box. Deduced amino acid sequences were obtained from D. rerio (GenBank accession no. NM_131047) and H. sapiens (NM_004343). (B) The predicted peptide features of crt. SP, signal peptide.
Mentions: In the present study, by RT-PCR, 3′- and 5′-RACE PCR, we obtained the full-length cDNA sequences of grp78, crt, perk, ire-1α, and atf-6α genes, which were shown to be 2544 bp, 1678 bp, 3776 bp, 3440 bp, and 2785 bp, respectively (Table 4). Sequencing revealed that their cDNAs included an ORF of 1956 bp, 1254 bp, 3240 bp, 3113 bp, and 1971 bp, respectively, encoding a protein of 652 amino acids, 418 amino acids, 1080 amino acids, 1038 amino acids, and 657 amino acids, respectively. The protein sequence of P. fulvidraco grp78 possessed all the characteristic features of grp78, including signal peptide, C-terminal KDEL motif, ATPase domain, and substrate-binding domain (Figure 1). The protein sequence of yellow catfish crt possessed all the characteristic features of crt, including signal peptide, N-glycosylation site, and C-terminal KDEL motif (Figure 2). The predicted amino acid sequence for the yellow catfish perk, ire-1α, and atf-6α possessed all the characteristic features of ER stress–sensing genes, including signal peptide, transmembrane domain, protein kinase, ribonuclease, sensor domain, and effector domain (Figure 3, Figure 4, and Figure 5).

Bottom Line: The predicted amino acid sequences for the yellow catfish grp78, crt, perk, ire-1α, and atf-6α revealed that the proteins contained all of the structural features that were characteristic of the five genes in other species, including the KDEL motif, signal peptide, sensor domain, and effector domain. mRNAs of the five genes mentioned above were expressed in various tissues, but their mRNA levels varied among tissues.For the first time, our study cloned grp78, crt, perk, ire-1α, and atf-6α genes in yellow catfish and demonstrated their differential expression among tissues.Moreover, the present study also indicated differential regulation of these ER stress-related genes by dietary Cu deficiency and excess, which will be beneficial for us to evaluate effects of dietary Cu levels in fish at the molecular level, based on the upstream pathway of lipid metabolism (the ER) and thus provide novel insights regarding the nutrition of Cu in fish.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Fishery College, Huazhong Agricultural University, Wuhan 430070, China Freshwater Aquaculture Collaborative Innovative Center of Hubei Province, Wuhan 430070, China.

No MeSH data available.