CDNA Cloning and Partial Characterization of the DJ-1 Gene from Tribolium castaneum.

Shunya Sasaki,Takuma Sakamoto,Michael R Kanost,Hiroko Tabunoki,Maaya Nishiko

doi:10.3390/antiox10121970

Abstract

The DJ-1 gene is highly conserved across a wide variety of organisms and it plays a role in anti-oxidative stress mechanisms in cells. The red flour beetle, Tribolium castaneum, is widely used as a model insect species because it is easy to evaluate gene function in this species using RNA interference (RNAi). The T. castaneum DJ-1 (TcDJ-1) sequence is annotated in the T. castaneum genome database; however, the function and characteristics of the TcDJ-1 gene have not been elucidated. Here, we investigated the cDNA sequence of TcDJ-1 and partially characterized its function. First, we examined the TcDJ-1 amino acid sequence and found that it was highly conserved with sequences from other species. TcDJ-1 mRNA expression was higher in the early pupal and adult developmental stages. We evaluated oxidant tolerance in TcDJ-1 knockdown adults using paraquat and found that adults with TcDJ-1 knockdown exhibited increased sensitivity to paraquat. Our findings show that TcDJ-1 has an antioxidant function, as observed for DJ-1 from other insects. Therefore, these results suggest that TcDJ-1 protects against oxidative stress during metamorphosis.

Highlights

We examined T. castaneum DJ-1 (TcDJ-1) gene-encoded amino acid residues corresponding to the Cys-His diad at positions 106 and 126 and found that the histidine at position 126 was replaced with tyrosine in TcDJ-1 (Table 1)
Our finding suggests that TcDJ-1 may play a role in protection from oxidative damage during metamorphosis, like DmDJ-1β
We found that TcDJ-1 likely has an antioxidant function similar to that of DJ-1 in D. melanogaster and B. mori DJ-1 and that TcDJ-1 knockdown does not affect

Summary

Introduction

The DJ-1 gene was discovered to be a proto-oncogene in cultured mouse cells [1], and it has been found to be a causative gene of Parkinson’s disease (PD), because mutations in the DJ-1 gene cause familial PD. The DJ-1 gene is highly conserved across animal species and is proposed to play important roles related to antioxidant function [4]. The human DJ-1 protein is multifunctional and plays roles in antioxidant ability, fertility, and protease activity [5,6,7,8]. One function of DJ-1 is to protect cells from oxidative damage to the mitochondria. Oxidative modification leads to mitochondrial damage in cultured cells exposed to several compounds, such as paraquat, which inhibits the electron transport chain of mitochondrial complex I [10]. DJ-1 seems to directly scavenge free radicals from mitochondria in response to oxidative stress

Methods

Results

Conclusion