title: Molecular cloning and characterization of 1-Cys and 2-Cys peroxiredoxins from the bumblebee Bombus ignitus.
abstract: We cloned and characterized two peroxiredoxins (Prxs), BiPrx1 (a 1-Cys Prx) and BiTPx1 (a 2-Cys Prx) from the bumblebee Bombus ignitus. The BiPrx1 gene consists of 5 exons, encoding 220 amino acid residues with one conserved cysteine residue. The BiTPx1 gene consists of three exons, encoding 195 amino acid residues with 2 conserved cysteine residues. Recombinant BiPrx1 (27 kDa) and BiTPx1 (25 kDa), expressed in baculovirus-infected insect Sf9 cells, reduced H2O2 in the presence of electrons donated by dithiothreitol. Unlike BiTPx1, however, BiPrx1 did not show reduction activity when thioredoxin was used as the electron donor. Both BiPrx1 and BiTPx1 protected super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Tissue distribution analyses showed the presence of BiPrx1 and BiTPx1 in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting that BiPrx1 and BiTPx1 are not secretable. When H2O2 was injected into B. ignitus bees, BiPrx1 and BiTPx1 transcripts were acutely up-regulated in the fat body tissues. We also demonstrated the regulation of BiPrx1 and BiTPx1 expression via reduction of transcript levels in the fat body with RNA interference (RNAi). Under H2O2 overload, the RNAi-induced BiPrx1 knock-down B. ignitus worker bees showed up-regulated expression of BiTPx1. Reciprocally, BiTPx1 RNAi knockdowns showed up-regulated BiPrx1 expression in the fat body. These results indicate that the loss of expression of BiPrx1 or BiTPx1 is compensated by the up-regulation of expression of the other peroxidase in response to H2O2 overload.
authors.affiliation: College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Republic of Korea.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Hu
authors.forename: Kwang Sik
authors.initials: KS
authors.lastname: Lee
authors.forename: Young Moo
authors.initials: YM
authors.lastname: Choo
authors.forename: Hyung Joo
authors.initials: HJ
authors.lastname: Yoon
authors.forename: Sang Mong
authors.initials: SM
authors.lastname: Lee
authors.forename: Jai Heon
authors.initials: JH
authors.lastname: Lee
authors.forename: Doh Hoon
authors.initials: DH
authors.lastname: Kim
authors.forename: Hung Dae
authors.initials: HD
authors.lastname: Sohn
authors.forename: Byung Rae
authors.initials: BR
authors.lastname: Jin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001516:Bees / Q000033:anatomy & histology / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D005216:Fat Body / Q000378:metabolism; D020869:Gene Expression Profiling; D015723:Gene Library; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008969:Molecular Sequence Data; D054464:Peroxiredoxins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D054817:Pollination; D034622:RNA Interference; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins