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Superoxide dismutases, SOD1 and SOD2, play a distinct role in the fat body during pupation in silkworm Bombyx mori.

Nojima Y, Ito K, Ono H, Nakazato T, Bono H, Yokoyama T, Sato R, Suetsugu Y, Nakamura Y, Yamamoto K, Satoh J, Tabunoki H, Fugo H - PLoS ONE (2015)

Bottom Line: One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria.We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage.These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183-8509, Japan; Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Noshio, 2-522-1, Kiyose, Tokyo 204-8588, Japan.

ABSTRACT
One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria. We characterized SOD1 and SOD2 of Bombyx mori isolated from the fat body of larvae. Immunological analysis demonstrated the presence of BmSOD1 and BmSOD2 in the silk gland, midgut, fat body, Malpighian tubules, testis and ovary from larvae to adults. We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage. The anti-oxidative functions of BmSOD1 and BmSOD2 were assessed by exposing larvae to insecticide rotenone or vasodilator isosorbide dinitrate, which is an ROS generator in BmN4 cells; however, exposure to these compounds had no effect on the expression levels of either BmSOD protein. Next, we investigated the physiological role of BmSOD1 and BmSOD2 under environmental oxidative stress, applied through whole-body UV irradiation and assayed using quantitative RT-PCR, immunoblotting and microarray analysis. The mRNA expression level of both BmSOD1 and BmSOD2 was markedly increased but protein expression level was increased only slightly. To examine the differences in mRNA and protein level due to UV irradiation intensity, we performed microarray analysis. Gene set enrichment analysis revealed that genes in the insulin signaling pathway and PPAR signaling pathway were significantly up-regulated after 6 and 12 hours of UV irradiation. Taken together, the activities of BmSOD1 and BmSOD2 may be related to the response to UV irradiation stress in B. mori. These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

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Amino acid alignment and tree of SOD1 and SOD2 from B. mori and other species.Amino acid sequence alignment of SOD1 (A) and SOD2 (B) of B. mori and other species: Homo sapiens, Rattus norvegicus, Mus musculus, Danio rerio, Xenopus tropicalis, Anopheles gambiae, Drosophila melanogaster, Plutella xylostella (a; PXGS_V2_006641, b; PXGS_V2_031456), and Manduca sexta. Conserved amino acids H (His), E (Glu) and D (Asp) of human SOD1 and SOD2 are marked by red asterisks (*). Levels of conserved amino acid residues among the various species are graphically shown below the sequences. Residues in the alignment are color-coded according to the Rasmol color scheme (http://life.nthu.edu.tw/~fmhsu/rasframe/COLORS.HTM#aminocolors) (C) Cluster tree for SOD1 and SOD2 of B. mori and other species.
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pone.0116007.g001: Amino acid alignment and tree of SOD1 and SOD2 from B. mori and other species.Amino acid sequence alignment of SOD1 (A) and SOD2 (B) of B. mori and other species: Homo sapiens, Rattus norvegicus, Mus musculus, Danio rerio, Xenopus tropicalis, Anopheles gambiae, Drosophila melanogaster, Plutella xylostella (a; PXGS_V2_006641, b; PXGS_V2_031456), and Manduca sexta. Conserved amino acids H (His), E (Glu) and D (Asp) of human SOD1 and SOD2 are marked by red asterisks (*). Levels of conserved amino acid residues among the various species are graphically shown below the sequences. Residues in the alignment are color-coded according to the Rasmol color scheme (http://life.nthu.edu.tw/~fmhsu/rasframe/COLORS.HTM#aminocolors) (C) Cluster tree for SOD1 and SOD2 of B. mori and other species.

Mentions: Alignment of the deduced BmSOD1 and BmSOD2 amino acid sequences and SOD1 and SOD2 orthologs from other species showed that the BmSOD1 protein sequence contains all of the conserved His and Asp residues (Fig. 1-A, red asterisks). In addition, the BmSOD2 protein sequence contains all of the conserved His, Asp and Glu residues (Fig. 1-B, red asterisks). These amino acid residues are involved in the coordination of the metal domain.


Superoxide dismutases, SOD1 and SOD2, play a distinct role in the fat body during pupation in silkworm Bombyx mori.

Nojima Y, Ito K, Ono H, Nakazato T, Bono H, Yokoyama T, Sato R, Suetsugu Y, Nakamura Y, Yamamoto K, Satoh J, Tabunoki H, Fugo H - PLoS ONE (2015)

Amino acid alignment and tree of SOD1 and SOD2 from B. mori and other species.Amino acid sequence alignment of SOD1 (A) and SOD2 (B) of B. mori and other species: Homo sapiens, Rattus norvegicus, Mus musculus, Danio rerio, Xenopus tropicalis, Anopheles gambiae, Drosophila melanogaster, Plutella xylostella (a; PXGS_V2_006641, b; PXGS_V2_031456), and Manduca sexta. Conserved amino acids H (His), E (Glu) and D (Asp) of human SOD1 and SOD2 are marked by red asterisks (*). Levels of conserved amino acid residues among the various species are graphically shown below the sequences. Residues in the alignment are color-coded according to the Rasmol color scheme (http://life.nthu.edu.tw/~fmhsu/rasframe/COLORS.HTM#aminocolors) (C) Cluster tree for SOD1 and SOD2 of B. mori and other species.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4340916&req=5

pone.0116007.g001: Amino acid alignment and tree of SOD1 and SOD2 from B. mori and other species.Amino acid sequence alignment of SOD1 (A) and SOD2 (B) of B. mori and other species: Homo sapiens, Rattus norvegicus, Mus musculus, Danio rerio, Xenopus tropicalis, Anopheles gambiae, Drosophila melanogaster, Plutella xylostella (a; PXGS_V2_006641, b; PXGS_V2_031456), and Manduca sexta. Conserved amino acids H (His), E (Glu) and D (Asp) of human SOD1 and SOD2 are marked by red asterisks (*). Levels of conserved amino acid residues among the various species are graphically shown below the sequences. Residues in the alignment are color-coded according to the Rasmol color scheme (http://life.nthu.edu.tw/~fmhsu/rasframe/COLORS.HTM#aminocolors) (C) Cluster tree for SOD1 and SOD2 of B. mori and other species.
Mentions: Alignment of the deduced BmSOD1 and BmSOD2 amino acid sequences and SOD1 and SOD2 orthologs from other species showed that the BmSOD1 protein sequence contains all of the conserved His and Asp residues (Fig. 1-A, red asterisks). In addition, the BmSOD2 protein sequence contains all of the conserved His, Asp and Glu residues (Fig. 1-B, red asterisks). These amino acid residues are involved in the coordination of the metal domain.

Bottom Line: One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria.We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage.These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183-8509, Japan; Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Noshio, 2-522-1, Kiyose, Tokyo 204-8588, Japan.

ABSTRACT
One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria. We characterized SOD1 and SOD2 of Bombyx mori isolated from the fat body of larvae. Immunological analysis demonstrated the presence of BmSOD1 and BmSOD2 in the silk gland, midgut, fat body, Malpighian tubules, testis and ovary from larvae to adults. We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage. The anti-oxidative functions of BmSOD1 and BmSOD2 were assessed by exposing larvae to insecticide rotenone or vasodilator isosorbide dinitrate, which is an ROS generator in BmN4 cells; however, exposure to these compounds had no effect on the expression levels of either BmSOD protein. Next, we investigated the physiological role of BmSOD1 and BmSOD2 under environmental oxidative stress, applied through whole-body UV irradiation and assayed using quantitative RT-PCR, immunoblotting and microarray analysis. The mRNA expression level of both BmSOD1 and BmSOD2 was markedly increased but protein expression level was increased only slightly. To examine the differences in mRNA and protein level due to UV irradiation intensity, we performed microarray analysis. Gene set enrichment analysis revealed that genes in the insulin signaling pathway and PPAR signaling pathway were significantly up-regulated after 6 and 12 hours of UV irradiation. Taken together, the activities of BmSOD1 and BmSOD2 may be related to the response to UV irradiation stress in B. mori. These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

Show MeSH
Related in: MedlinePlus