Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model
Abstract
:1. Introduction
2. Results
2.1. Cloning and Sequence Analysis of zip10, zip13, and zip14 Promoters of Yellow Catfish
2.2. 5′-Sequence Deletion Mutation Assays of zip10, zip13, and zip14 Promoters of Yellow Catfish
2.3. Site-Mutation Assays of the MTF-1, KLF4, and STAT3 Binding Sites in the zip10, zip13 and zip14 Promoters of Yellow Catfish
2.4. Electrophoretic Mobility Shift Assay
2.5. Response of pfZip10-EGFP, pfZip13-EGFP, and pfZip14-EGFP to Zn Administration in HEK293T Cells
2.6. The Subcellular Localization of the Zip10, Zip13, and Zip14 Proteins of Yellow Catfish in HEK293T Cells
3. Discussion
4. Materials and Methods
4.1. Animals, Cells, and Reagents
4.2. Promoters Cloning and Plasmids Construction
4.3. Promoter Sequence Analysis
4.4. Dual Luciferase Activities Assays
4.5. Site Mutation Analysis of TFBS in the zip10, zip13, and zip14 Promoters
4.6. Electrophoretic Mobility-Shift Assay
4.7. Fluorescence Intensity Analysis
4.8. Subcellular Localization Investigation of Zip10, Zip13, and Zip14
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
References
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Name | Location in zip10 Genomic Locus | Location in zip13 Genomic Locus | Location in zip14 Genomic Locus |
---|---|---|---|
MRE | −2017 bp/−2004 bp | −360 bp/−345 bp | −1457 bp/−1442 bp |
PPARα:RXR | −1997 bp/−1980 bp | −543 bp/−529 bp | None |
FOXO4 | −1976 bp/−1970 bp | −981 bp/−975 bp | −1687 bp/−1681 bp |
FOXO6 | −1043 bp/−1037 bp | −248 bp/−242 bp | −681 bp /−675 bp and −435 bp /−429 bp |
SREBP | −1760 bp/−1751 bp | None | −1785 bp/−1776 bp |
CREB1 | −1729 bp/−1718 bp and −1330 bp/−1319 bp | None | −1293 bp/−1286 bp and −119 bp/−112 bp |
AP1 | None | −1393 bp/−1383 bp | None |
ZEB1 | −1601 bp/−1596 bp | −651/−640 bp | −1306 bp /−1301 bp |
RREB1 | −353 bp/−334 bp | −731 bp/−724 bp | None |
SP1 | −1423 bp/−1414 bp | None | −1253 bp/−1243 bp |
KLF4 | −1124 bp/−1114 bp | −606 bp/−594 bp | None |
STAT3 | None | None | (−1383 bp/−1375 bp and −1358 bp/−1348 bp) |
STAT5a:STAT5b | −456 bp/−445 bp | None | None |
STAT6 | −960 bp/−946 bp | −1126 bp/−1212 bp | None |
CEBPα | −890 bp/−881 bp and −907 bp/−899 bp | −1300 bp/−1292 bp and −1123 bp/−1115 bp | −206 bp/−196 bp |
ATF4 | None | −1027 bp/−1014 bp | None |
CCAAT-box | −687 bp/−676 bp | −106/−95 bp | None |
TATA-box | −319 bp/−305 bp | −92 bp/−78 bp | None |
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Liu, S.-Z.; Xu, Y.-C.; Tan, X.-Y.; Zhao, T.; Zhang, D.-G.; Yang, H.; Luo, Z. Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model. Int. J. Mol. Sci. 2022, 23, 8034. https://doi.org/10.3390/ijms23148034
Liu S-Z, Xu Y-C, Tan X-Y, Zhao T, Zhang D-G, Yang H, Luo Z. Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model. International Journal of Molecular Sciences. 2022; 23(14):8034. https://doi.org/10.3390/ijms23148034
Chicago/Turabian StyleLiu, Sheng-Zan, Yi-Chuang Xu, Xiao-Ying Tan, Tao Zhao, Dian-Guang Zhang, Hong Yang, and Zhi Luo. 2022. "Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model" International Journal of Molecular Sciences 23, no. 14: 8034. https://doi.org/10.3390/ijms23148034