Molecular cloning and characterization of peroxiredoxin 4 involved in protection against oxidative stress in the silkworm Bombyx mori

Abstract

Peroxiredoxins (Prxs) are a ubiquitous family of proteins that play important roles in insects in protection against oxidative stress through the detoxification of cellular peroxides. Here, we describe the cloning and characterization of a Prx4 cDNA of the silkworm Bombyx mori (BmPrx4). The BmPrx4 gene has an open reading frame of 744 bp encoding 248 amino acids and a conserved motif, VCP, involved in its presumed redox functions. The heterologously expressed proteins of the gene in Escherichia coli showed antioxidant activity, removed hydrogen peroxide and protected DnA. Western blotting analysis showed the presence of BmPrx4 in the haemolymph, suggesting that the protein is secretable. Moreover, BmPrx4 was expressed at all developmental stages. The expression level of BmPrx4 was relatively low during the feeding stage but high at the wandering stage. BmPrx4 was induced by quercetin or temperature stress. Immunohistochemical analysis revealed that BmPrx4 is present in the brain, neurones and olfactory organ of the head in silkworms. Overall, our results indicate that the expression profile of BmPrx4 correlates well with protection from oxidative damage. Our data provide clues for the development of control technology for agricultural and forestry pests as the silkworm is a representative of lepidopteran pests.

Comparative analyses of salivary proteins from three aphid species

“…Hydrogen per-oxide can also kill pathogens directly and strengthen plant cell walls by oxidative cross-linking of cell wall structural proteins. In many pathogens and insects, peroxiredoxins are peroxide-detoxifying enzymes that play important roles in protection against oxidative stress (Piacenza et al, 2008;Dubreuil et al, 2011;Shi et al, 2012). Although a peroxidase has been identified in the saliva of the grain aphid Sitobion avenae (Rao et al, 2013), and peroxidase activity has been measured in the salivary secretions of several aphid species (Cherqui & Tjallingii, 2000), the present study is the first to report the identification of peroxiredoxins in aphid saliva.…”

A novel 1-Cys thioredoxin peroxidase gene in Apis cerana cerana: characterization of AccTpx4 and its role in oxidative stresses

“…The recombinant protein existed mostly as a soluble protein (Fig. 7a), the same as BmPrx4 (Shi et al 2012). Furthermore, the purified recombinant AccTpx4 protein could efficiently degrade H 2 O 2 in the presence of DTT (Fig.…”

“…Prxs are induced by various stimuli such as temperature stress (Shi et al 2012;Wang et al 2008) and multiple other environmental stresses (Yu et al 2011). To characterize the transcript levels of AccTpx4 under various environmental stresses, adult bees were exposed to an artificial hostile environment or were fed reagents that cause oxidative stress and then Q-PCR was performed.…”

First Report of a Peroxiredoxin Homologue in Jellyfish: Molecular Cloning, Expression and Functional Characterization of CcPrx4 from Cyanea capillata

“…In the assay for in vitro peroxidase activity, the recombinant CcPrx4 protein could scavenge H 2 O 2 and exhibited a concentration-dependent peroxidase activity, quickly scavenging H 2 O 2 in about two minutes. The MCO system has been widely used to assess ROS damage to DNA and has been previously used to assay the antioxidant activity of Prx proteins from different subfamilies [62,63,64]. In this study, we found that the CcPrx4 protein could protect plasmid DNA from nicking.…”