Title | Biomphalaria glabrata peroxiredoxin: Effect of Schistosoma mansoni infection on differential gene regulation |
Publication Type | Journal Article |
Year of Publication | 2009 |
Authors | Knight, M, Raghavan, N, Goodall, C, Cousin, C, Ittiprasert, W, Sayed, A, Miller, A, Williams, DL, Bayne, CJ |
Journal | Molecular and Biochemical Parasitology |
Volume | 167 |
Pagination | 20-31 |
Type of Article | Journal Article |
ISSN | 0166-6851 |
Abstract | To identify gene(s) that may be associated with resistance/susceptibility in the intermediate snail host Biomphalaria glabrata to Schistosoma mansoni infection, a snail albumen gland cDNA library was differentially screened and a partial cDNA encoding an antioxidant enzyme thioredoxin peroxidase (Tpx), or peroxiredoxin (Prx), was identified. The 753 bp full-length, single-copy, constitutively expressed gene now referred to as BgPrx4 was later isolated. BgPrx4 is a 2-Cys peroxiredoxin containing the conserved peroxidatic cysteine (C(P)) in the N-terminus and the resolving cysteine (C(R)) in the C-terminus. Sequence analysis of BgPrx4 from both resistant and susceptible snails revealed the presence of several (at least 7) single nucleotide polymorphisms (SNPs). Phylogenetic analysis indicated BgPrx4 to resemble a homolog of human peroxiredoxin, PRDX4. Northern analysis of hepatopancreas RNA from both resistant and susceptible snails showed that upon parasite exposure there were qualitative changes in gene expression. Quantitative real-time RT-PCR analysis showed differences in the levels of BgPrx4 transcript induction following infection, with the transcript up-regulated in resistant snails during the early phase (5 h) of infection compared to susceptible snails in which it was down-regulated within the early time period. While there was an increase in transcription in susceptible snails later (48 h) post-infection, this never reached the levels detected in resistant snails. A similar trend - higher, earlier up-regulation in the resistant snails but lower, slower protein expression in susceptible snails - was observed by Western blot analysis. Enzymatic analysis of the purified, recombinant BgPrx4 revealed the snail sequence to function as PT-x but with an unusual ability to use both thioredoxin and glutathione as substrates. (C) 2009 Elsevier B.V. All rights reserved. |
URL | <Go to ISI>://WOS:000268049600003 |
DOI | 10.1016/j.molbiopara.2009.04.002 |