Molecular cloning and characterization of ATX1 cDNA from the mole cricket, Gryllotalpa orientalis.
To search for an insect homologue of antioxidant protein 1 (ATX1), a mole cricket, Gryllotalpa orientalis, cDNA library was screened and a cDNA clone, which encodes a 73 amino acid polypeptide with a predicted molecular mass of 8.0 kDa and pI of 5.68, was isolated. The G. orientalis ATX1 (GoATX1) cDNA features both a MTCXXC copper-binding site in the N-terminus and a KTGK lysine-rich region in the C-terminus. The deduced amino acid sequence of the GoATX1 cDNA showed 63% identity to Drosophila melanogaster ATX1 and 55% to Ixodes pacificus ATX1. Northern blot analysis revealed the presence of GoATX1 transcripts in midgut, fat body, and epidermis. When H2O2 was injected into the body cavity of G. orientalis adult, GoATX1 mRNA expression was up-regulated in the fat body tissue. Fat body expression level of GoATX1 mRNA in the fat body was increased following exposure to low (4 degrees C) and high (37 degrees C) temperatures, suggesting that GoATX1 plays a protective role against oxidative stress caused by temperature shock. This is the first report about a functional role of insect ATX1 in antioxidant defense.
Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA, Suwon, Korea. ikkim81@rda.go.kr
Iksoo
I
Kim
Kwang Sik
KS
Lee
Jae Sam
JS
Hwang
Mi Young
MY
Ahn
Eun Young
EY
Yun
Jian Hong
JH
Li
Hung Dae
HD
Sohn
Byung Rae
BR
Jin
D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003300:Copper / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006135:Gryllidae / Q000235:genetics* / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D015854:Up-Regulation
D000975:Antioxidants; D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D003300:Copper; D006861:Hydrogen Peroxide