Sequence, Expression, and Anti-GCRV Function of the Ferritin from the Grass Carp, <i>Ctenopharyngodon idellus</i>
Ferritin possesses an immune function to defend against pathogen infection. To elucidate the immunity-protecting roles of ferritin from <i>Ctenopharyngodon idellus</i> (<i>Ciferritin</i>) against virus infection, the cDNA and promoter sequences of <i>Ciferritin</i>...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-06-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/12/6835 |
| Summary: | Ferritin possesses an immune function to defend against pathogen infection. To elucidate the immunity-protecting roles of ferritin from <i>Ctenopharyngodon idellus</i> (<i>Ciferritin</i>) against virus infection, the cDNA and promoter sequences of <i>Ciferritin</i> were determined, and the correlations between <i>Ciferrtin</i> expressions and promoter methylation levels were analyzed. In addition, the functional role of Ciferrtin on GCRV (grass carp reovirus) infection was assessed. The full-length cDNA of <i>Ciferritin</i> is 1053 bp, consists of a 531 bp open-reading frame, and encodes 176 amino acids. Ciferritin showed the highest sequence identity with the ferritin middle subunit of <i>Mylopharyngodon piceus</i> (93.56%), followed by the subunits of <i>Megalobrama amblycephala</i> and <i>Sinocyclocheilus rhinocerous</i>. Ciferritin contains a conserved ferritin domain (interval: 10–94 aa), and the caspase recruitment domain (CARD) and Rubrerythrin domain were also predicted. In the spleen and kidney, significantly higher <i>Ciferritin</i> expressions were observed at 6, 12, 24, or 168 h post GCRV infection than those in the PBS injection group (<i>p</i> < 0.05). The <i>Ciferrtin</i> expression level in the progeny of maternal-immunized grass carp was significantly higher than that in the progeny of common grass carp (<i>p</i> < 0.05). <i>Ciferritin</i> promoter methylation level in the progeny from common grass carp was 1.27 ± 0.15, and in the progeny of the maternal-immunized group was 1.00 ± 0.14. In addition, methylation levels of “CpG9” and “CpG10” loci were significantly lower in the progeny of maternal-immunized fish than those in the common group. Except for the “CpG5”, methylation levels of all other detected “CpG” loci negatively correlated with <i>Ciferritin</i> expression levels. Furthermore, the total methylation level of “CpG1–10” negatively correlated with the <i>Ciferritin</i> expressions. The <i>Ciferritin</i> expression level was significantly up-regulated, and the VP7 protein levels were significantly reduced, at 24 h post GCRV infection in the <i>Ciferritin</i> over-expression cells (<i>p</i> < 0.05). The results from the present study provide sequence, epigenetic modification and expression, and anti-GCRV functional information of Ciferritin, which provide a basis for achieving resistance to GCRV in grass carp breeding. |
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| ISSN: | 1661-6596 1422-0067 |