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Tracing and exploring the evolutionary origin and systematic function of fish complement C9

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Abstract

Complement C9, as a member of terminal complement component (TCC) protein, plays important roles in innate immunity. However, some complement components appear to show difference and evolutionary complexity between higher and lower vertebrates. Hence, it is essential to carry on a study of evolutionary origin and systematic function of C9 in fish and non-fish vertebrates. This study aims to explore the complement gene evolution and potential function in fish based on molecular and structural biology. Herein, we found complete divergence of C9 throughout the gene evolution. The optimal codons of C9 sequences tended to be closer to the genomes of lower vertebrates compared to higher vertebrates. Further, conserved amino acids in the C9 TMH1 region were identified, implying their potential functional association with MAC growth and pore formation. Transposons and simple repeats, as gene elements, exhibited a differential distribution in the genomic regions in different animal groups but were sparsely scattered around the sixth exon (TMH1 region). Notably, this demonstrated the regulatory complexity of the C9 gene in higher vertebrates. The negative selection pressures on fish and non-fish groups improved both the sequence conservation and similarity. Through gene/protein regulatory network and pathway analyses, the systematic function of C9 protein was showcased; thus, we could reveal the divergence of the systematic function of C9 across species from different evolutionary positions. In addition, more complicated functions of C9 in higher vertebrates could established by the altered spatial conformation of the protein. Collectively, the present study illustrates the C9 gene evolutionary process and the difference in its systematic function across multiple species. Such advances provide new insights for understanding the evolutionary and potential functions of complement C9.

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Acknowledgements

We thank FreeScience Information Technology Company (Zhejiang province, China) for proofreading, and editing the grammar in the text.

Funding

This study was funded by the China’s Agricultural Research System (CARS-45-03) and the Project of Shanghai Engineering and Technology Center for Promoting Ability (16DZ2281200).

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  1. Lisen Li and Yubang Shen contributed equally to the article.

Authors and Affiliations

  1. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, College of Aquaculture and Life Science, Shanghai Ocean University, Shanghai, 201306, China

    Lisen Li, Yubang Shen, Xiaoyan Xu, Weining Yang & Jiale Li

  2. Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, 201306, China

    Jiale Li

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  1. Lisen Li
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  2. Yubang Shen
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Correspondence to Yubang Shen or Jiale Li.

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Supplementary Information

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438_2021_1773_MOESM1_ESM.pdf

Supplementary file1Supplementary Fig. 1. Multiple sequence alignment of C9. The predicted signal peptide is underlined at the beginning of the amino acid sequence. Amino acids are shaded according to physiochemical properties: Proline Blue/Red; Glycine Green/Red; Tiny Blue/Yellow; Small Green/Yellow; Positive Red/Blue; Negative Green/Blue; Charged White/Blue; Amphoteric Red/Green; Polar Black/Green; Aliphatic Red/Gray; Aromatic Blue/Gray; Hydrophobic White/Black). Complete residue conservation is annotated with a ‘‘*” and highlighted in bold. Partially conserved residues are denoted by ‘‘.’’ and the position of the consensus amino acids present in the greatest number of sequences are indicated with ‘‘:”. Gaps inside the sequence are marked by dashes. TMH1 region is framed by the red dashed box. MACPF domain is denoted by a black line on the top of the sequence. The calcium-binding sites are marked by the black arrow (PDF 560 KB)

438_2021_1773_MOESM2_ESM.xlsx

Supplementary file2Supplementary Table 1. Accession numbers of different species C9 amino acid sequences used in the analysis (XLSX 13 KB)

Supplementary file3Supplementary Table 2. Information of sites/residues under selective pressure (XLSX 10 KB)

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Li, L., Shen, Y., Xu, X. et al. Tracing and exploring the evolutionary origin and systematic function of fish complement C9. Mol Genet Genomics 296, 665–676 (2021). https://doi.org/10.1007/s00438-021-01773-2

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