title: BmDJ-1 is a key regulator of oxidative modification in the development of the silkworm, Bombyx mori.
abstract: We cloned cDNA for the Bombyx mori DJ-1 protein (BmDJ-1) from the brains of larvae. BmDJ-1 is composed of 190 amino acids and encoded by 672 nucleotides. Northern blot analysis showed that BmDJ-1 is transcribed as a 756-bp mRNA and has one isoform. Reverse transcriptase (RT)-PCR experiments revealed that the BmDJ-1 was present in the brain, fatbody, Malpighian tubule, ovary and testis but present in only low amounts in the silkgland and hemocyte of day 4 fifth instar larvae. Immunological analysis demonstrated the presence of BmDJ-1 in the brain, midgut, fatbody, Malpighian tubule, testis and ovary from the larvae to the adult. We found that BmDJ-1 has a unique expression pattern through the fifth instar larval to adult developmental stage. We assessed the anti-oxidative function of BmDJ-1 using rotenone (ROT) in day 3 fifth instar larvae. Administration of ROT to day 3 fifth instar larvae, together with exogenous (BmNPV-BmDJ-1 infection for 4 days in advance) BmDJ-1, produced significantly lower 24-h mortality in BmDJ-1 groups than in the control. 2D-PAGE revealed an isoelectric point (pI) shift to an acidic form for BmDJ-1 in BmN4 cells upon ROT stimulus. Among the factors examined for their effects on expression level of BmDJ-1 in the hemolymph, nitric oxide (NO) concentration was identified based on dramatic developmental stage-dependent changes. Administration of isosorbide dinitrate (ISDN), which is an NO donor, to BmN4 cells produced increased expression of BmDJ-1 compared to the control. These results suggest that BmDJ-1 might control oxidative stress in the cell due to NO and serves as a development modulation factor in B. mori.
authors: Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Tokyo, Japan. tabunoki@my-pharm.ac.jp
authors: Hiroko
authors: H
authors: Tabunoki
authors: Hiroaki
authors: H
authors: Ode
authors: Yutaka
authors: Y
authors: Banno
authors: Susumu
authors: S
authors: Katsuma
authors: Toru
authors: T
authors: Shimada
authors: Kazuei
authors: K
authors: Mita
authors: Kimiko
authors: K
authors: Yamamoto
authors: Ryoichi
authors: R
authors: Sato
authors: Reiko
authors: R
authors: Ishii-Nozawa
authors: Jun-ichi
authors: J
authors: Satoh
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D012831:Bombyx / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006458:Hemolymph / Q000187:drug effects / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012402:Rotenone / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D019476:Insect Proteins; D012402:Rotenone