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Molecular cloning and functional characterization of the hypoxia-inducible factor-1α in bighead carp (Aristichthys nobilis)

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Abstract

HIF-l is the earliest documented and most widely studied hypoxia-inducible factor (HIF) and plays a key role in the cell hypoxia signal transduction pathway. Particularly, the HIF-1α protein is sensitive to oxygen and plays a critical role in hypoxia regulation. This study is the first to report on the molecular cloning and characterization of HIF-1α in bighead carp (Aristichthys nobilis; anHIF-1α). The full-length cDNA of anHIF-1α was 2361 bp, and encodes an estimated 674 amino acids with a predicted molecular mass of 76.10 kDa and a theoretical isoelectric point of 7.72. Moreover, the conserved basic Helix-Loop-Helix domain along with two Per-ARNT-Sim domains (A/B), and C-TAD were identified in this protein. Interestingly, the tertiary structure of the anHIF-1α protein was found to be extremely similar to that of mice. Multiple comparison and phylogenetic tree results demonstrated that anHIF-1α was highly conserved. Under normoxic conditions, anHIF-1α mRNA transcripts could be detected in all tissues examined with the highest expression level in the heart. With gradually decreasing oxygen concentrations, anHIF-1α mRNA level was upregulated significantly in the gill, liver, kidney, spleen, intestine, brain, and muscle tissues (P < 0.05). Similarly, anHIF-1α was expressed in all examined bighead carp tissues, and the results suggested that the upregulation of anHIF-1α at the transcriptional level may be an important stress response adaptation to hypoxia in bighead carp. Finally, based on the tertiary structure comparative analyses between anHIF-1α with mouse HIF-1α, we think the physiological function, and protein structure of HIF-1α could be compared between fish and mammal in the future.

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Data availability

The data that support the findings of this study are available from the corresponding author Ge Xianping upon reasonable request at the Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences.

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Acknowledgments

We would like to thank the postgraduate students of Fish Disease and Nutrition Department, Freshwater Fisheries Research Center (FFRC), Chinese Academy of Fishery Sciences (CAFS), PR China, for their help throughout the research period.

Funding

This work was supported by the Earmarked Fund for China Agriculture Research System (Grant No. CARS-45), and Science and Technology Innovation Team (Grant No. 2020TD59).

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Authors and Affiliations

  1. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China

    Yan Lin, Ling-Hong Miao, Bo Liu, Bing-Wen Xi, Liang-Kun Pan & Xian-Ping Ge

  2. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, 214081, China

    Ling-Hong Miao, Bo Liu, Bing-Wen Xi & Xian-Ping Ge

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  1. Yan Lin
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  2. Ling-Hong Miao
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  4. Bing-Wen Xi
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  5. Liang-Kun Pan
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  6. Xian-Ping Ge
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Contributions

Yan Lin, Ling-Hong Miao, and Xian-Ping Ge conceived the idea and designed the project. Bo Liu, Bing-Wen Xi, and Liang-Kun Pan performed the experiments. Yan Lin analyzed the data and wrote the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Xian-Ping Ge.

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All experimental protocols were approved by the Bioethical Committee of Freshwater Fisheries Research Center (FFRC) of Chinese Academy of Fishery Sciences (CAFS) (BC 2013863, 9/2013)

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Lin, Y., Miao, LH., Liu, B. et al. Molecular cloning and functional characterization of the hypoxia-inducible factor-1α in bighead carp (Aristichthys nobilis). Fish Physiol Biochem 47, 351–364 (2021). https://doi.org/10.1007/s10695-020-00917-2

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