Parasitization by Macrocentrus cingulum (Hymenoptera: Braconidae) influences expression of prophenoloxidase in Asian corn borer Ostrinia furnacalis.

A prophenoloxidase (PPO) cDNA (OfPPO) was cloned from the Asian corn borer Ostrinia furnacalis. Sequence analysis revealed a full length transcript of the OfPPO cDNA with 2,686 bp, containing a 2,079 bp open reading frame (ORF), a 73-bp 5'-untranslated region, and a 534-bp 3'-untranslated region with a poly(A) signal. The ORF encodes a 693-amino acid polypeptide, containing two distinct copper-binding regions, a plausible thiol ester site, two proteolytic activation sites, and a conserved C-terminal region, but lacks a signal peptide sequence. Expression of the OfPPO transcript in the plasma, hemocytes, fat body and midgut was inhibited by Macrocentrus cingulum at 4 h post-parasitization (pp). In situ hybridization analysis showed that the hemocytes, especially the oenocytoids, hybridized strongly with the DNA probes of the OfPPO gene. No signal was detected in the cuticular epithelium or fat body of the parasitized larvae. Colloidal gold particles were used to visualize the PPO by immunoelectron microscopy. The time course study revealed a decrease in the labeling of the OfPPO at 4, 6, 8, 12, and 1 day pp in the larval integument and midgut parasitized by M. cingulum. We infer from time course studies of OfPPO gene expression and PO enzymatic activity that OfPPO in the integument is released from hemocytes and that the OfPPO expression was influenced at the transcriptional, translational, and then the post-translational level by parasitization challenge.

Molecular cloning and characterization of a novel human putative transmembrane protein homologous to mouse sideroflexin associated with sideroblastic anemia.

Sideroflexin1 (Sfxn1), the prototype of a novel family of evolutionarily conserved proteins present in eukaryotes, has been found mutated in mice with siderocytic anemia. It is speculated that this protein facilitates the transport of a component required for iron utilization into mitochondrial. During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA encoding a novel sideroflexin protein (SFXN4), which showed 59% identity and 71% similarity to mouse sideroflexin4. According to the search of the human genome database, SFXN4 gene is mapped to chromosome 10q25-26 and spans more than 24.7kb of the genomic DNA. It is 1428 base pair in length and the putative protein contains 305 amino acids with a conserved predicted five-transmembrane-domains structure. RT-PCR result shows that the SFXN4 gene is expressed in many tissues.