The Giardia Tom40 is capable of mediating its own import and Ginsenoside-Ro membrane insertion, given the highly simplified nature of the TOM complex in mitosomes. Our proteomics data support the hypothesis that ISC assembly is an important and possibly the only biosynthetic function of Giardia mitosomes. Previous phylogenic analyses have indicated that the ISC assembly machinery was obtained from the alphaproteobacterial endosymbiont; nearly complete ISC assembly machinery is present from trypanosomatids to higher eukaryotes. Therefore, the absence of certain components, such as IscA-1, Iba57, and Ind, in the mitosomal machines is apparently due to a secondary 
loss of specific target proteins. Noteworthy, we did not identify any proteins that would carry FeS clusters in Giardia mitosomes, except for components of the FeS cluster assembly machinery itself. It seems likely then that the main role of mitosomes could be to export preassembled FeS clusters, or other compounds that are essential for the biogenesis of FeS proteins, to other cellular compartments. In mitochondria, the export of these enigmatic compounds is dependent on the membrane ABC ”halftransporter” Atm1 and sulfhydryl oxidase Erv1. In the mitosome-enriched fraction, we identified four ABC half-transporters by mass spectrometry, and another candidate was predicted based on phyletic profiling of the G. intestinalis genome. However, compared to other Atm1 homologues, these candidates lack the x-loop with the conserved arginine, which is essential for known Atm1 transporters. No protein with homology to Erv1 was found by proteomics or by analysis of the Giardia genome. Another remaining question pertains to the source of ATP that is required for the multiple processes identified in mitosomes including FeS cluster assembly and export, organelle division, protein import and protein folding. In E. histolytica, it has been shown that a mitochondrial carrier Mechlorethamine hydrochloride family protein localizes to mitosomes and exchanges ATP and ADP across the inner membrane, effecting the import ATP into mitosomes. E. cuniculi mitosomes contain a distinct bacterial nucleotide transporter that may fulfill the same function. However, our proteomic analysis did not revealed a candidate nucleotide transporter in the mitosomes of Giardia leaving open the question of ATP acquisition. In conclusion, using iTRAQ-based mass spectrometry and bioinformatics we identified 139 candidate mitosomal proteins. Mitosomal localization was confirmed experimentally for 20 of 44 proteins tested, suggesting the complete mitosomal proteome of Giardia to be of the order of 50-100 proteins. Previous genome analyses failed to predict any of the novel mitosomal proteins identified here; only by combining quantitative mass spectrometry and bioinformatics were these novel proteins identified. The small proteome of the G. intestinalis mitosome indicates a marked reduction in mitochondrial metabolic activity and reduced requirements for organelle biogenesis. These do not mirror the reductions seen in the mitosomal proteome of Cryptosporidium, supporting the view that lineage-specific reductions produce organelles with distinct metabolic pathways and specific ”short-cut” pathways for biogenesis. Our findings provide new insight into aspects of mitochondrial evolution and the basis from which to begin reconstructing the details of precisely how these organelles are built and replicated to support Giardia growth and division. Dengue virus is a member of the family Flaviviridae, which comprises single stranded positive sense RNA viruses such as West Nile Virus, Japanese encephalitis virus, yellow fever virus, as well as the pestivirus bovine viral diarrhea virus and the hepacivirus, hepatitis C virus. Dengue virus infections are a significant global health concern. Approximately 100 million cases of dengue fever infections are reported annually, of which 250,000�C500,000 cases comprise the more severe and life-threatening dengue hemorrhagic fever. It is estimated that 2.5 billion people live in areas that are at risk for dengue outbreaks.