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Identification and characterization of a Macrobrachium nipponense ferritin subunit regulated by iron ion and pathogen
Shengming Sun1, Zhimin Gu2, Hongtuo Fu3
1Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China; Key Laboratory of Genetic Breeding and Aquaculture Biology of Freshwater Fishes, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Abstract:
Ferritin, a major iron storage protein in most living organisms, plays a crucial role in iron metabolism. In this study, the ferritin subunit MnFer was identified in the oriental river prawn (Macrobrachium nipponense) and functionally characterized. The full-length cDNA of MnFer is 999 bp in size with a 122-bp 5'-untranslated region (UTR), a 364-bp 3'-UTR and a 513-bp open reading frame that encodes a protein possessing 171 amino acids and a deduced molecular weight of 19.40 kDa. Prawn ferritin transcripts are expressed in muscle, heart, hepatopancreas, gill, hemocytes, ovary and testis. Quantitative real-time PCR revealed that the abundance of ferritin transcript was highest in the hepatopancreas followed by muscle. Ferritin transcript expression in muscle increased six-fold 3 h after the injection of iron. In the gill, a four-fold increase in ferritin transcript expression was detected 3 h post-injection; the expression remained elevated for 48 h. Heart ferritin mRNA expression increased up to seven-fold at 24 h post-injection. No significant difference was found in the hepatopancreas. The iron binding capacity of recombinant ferritin protein was also demonstrated in this study. In hemocyte experiments, the transcriptional expression of MnFer showed the strongest response to Aeromonas hydrophila. As a whole, our study suggested that the ferritin from M. nipponense may play critical roles in cellular and organismic iron homeostasis along with in innate immune defense.
Insights
This study identifies and characterizes MnFer, a ferritin subunit in the oriental river prawn (Macrobrachium nipponense). MnFer plays key roles in iron metabolism and innate immunity, responding to iron and bacterial challenges.
Area of Science:
- * Molecular Biology
- * Biochemistry
- * Aquaculture
Background:
- * Ferritin is a vital iron storage protein essential for iron metabolism in living organisms.
- * Understanding ferritin function in invertebrates like the oriental river prawn (Macrobrachium nipponense) is crucial for their physiology and health.
Purpose of the Study:
- * To identify and functionally characterize the ferritin subunit MnFer in Macrobrachium nipponense.
- * To investigate the expression patterns of MnFer under iron stimulation and immune challenge.
- * To assess the iron-binding capacity of the recombinant MnFer protein.
Main Methods:
- * cDNA cloning and sequencing to determine the full-length MnFer sequence.
- * Quantitative real-time PCR (qRT-PCR) to analyze MnFer transcript expression in various tissues and under different conditions.
- * Expression and purification of recombinant MnFer protein to test its iron-binding capacity.
- * Hemocyte experiments to evaluate MnFer response to bacterial infection.
Main Results:
- * The full-length cDNA of MnFer (999 bp) encodes a 171-amino acid protein with a molecular weight of 19.40 kDa.
- * MnFer transcripts are expressed in multiple tissues, with highest levels in hepatopancreas and muscle.
- * Iron injection significantly upregulated MnFer expression in muscle, gill, and heart tissues.
- * Recombinant MnFer protein demonstrated iron-binding capacity.
- * MnFer transcriptional expression was highest in response to Aeromonas hydrophila in hemocytes.
Conclusions:
- * The identified MnFer subunit in Macrobrachium nipponense is involved in iron homeostasis.
- * MnFer plays a significant role in the innate immune defense of the prawn.
- * This research provides insights into the dual function of ferritin in iron metabolism and immunity in aquaculture species.
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