Massively Parallel Sequencing and Analysis of the Necator americanus Transcriptome
Abstract
Background: The blood-feeding hookworm Necator americanus infects hundreds of millions of people worldwide. In order to elucidate fundamental molecular biological aspects of this hookworm, the transcriptome of the adult stage of Necator americanus was explored using next-generation sequencing and bioinformatic analyses. Methodology/Principal Findings: A total of 19,997 contigs were assembled from the sequence data; 6,771 of these contigs had known orthologues in the free-living nematode Caenorhabditis elegans, and most of them encoded proteins with WD40 repeats (10.6%), proteinase inhibitors (7.8%) or calcium-binding EF-hand proteins (6.7%). Bioinformatic analyses inferred that the C. elegans homologues are involved mainly in biological pathways linked to ribosome biogenesis (70%), oxidative phosphorylation (63%) and/or proteases (60%); most of these molecules were predicted to be involved in more than one biological pathway. Comparative analyses of the transcriptomes of N. americanus and the canine hookworm, Ancylostoma caninum, revealed qualitative and quantitative differences. For instance, proteinase inhibitors were inferred to be highly represented in the former species, whereas SCP/Tpx-1/Ag5/PR-1/Sc7 proteins (=SCP/TAPS or Ancylostoma-secreted proteins) were predominant in the latter. In N. americanus, essential molecules were predicted using a combination of orthology mapping and functional data available for C. elegans. Further analyses allowed the prioritization of 18 predicted drug targets which did not have homologues in the human host. These candidate targets were inferred to be linked to mitochondrial (e.g., processing proteins) or amino acid metabolism (e.g., asparagine t-RNA synthetase). Conclusions: This study has provided detailed insights into the transcriptome of the adult stage of N. americanus and examines similarities and differences between this species and A. caninum. Future efforts should focus on comparative transcriptomic and proteomic investigations of the other predominant human hookworm, A. duodenale, for both fundamental and applied purposes, including the prevalidation of anti-hookworm drug targets.
Additional Information
© 2010 Cantacessi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received January 11, 2010; Accepted March 25, 2010; Published May 11, 2010. Editor: Matty Knight, Biomedical Research Institute, United States of America. This research was supported by grants from the National Human Genome Research Institute (NHGRI) and National Institutes of Health (MM) and the Australian Research Council, the Australian Academy of Science and the Australian-American Fulbright Commission (RBG). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. CC is the grateful recipient of International Postgraduate Research Scholarship (IPRS) from the Australian Government and a fee-remission scholarship through the University of Melbourne as well as the Clunies Ross (2008) and Sue Newton (2009) awards from the School of Veterinary Science of the same university. The contribution of staff at WormBase is gratefully acknowledged. Author Contributions: Conceived and designed the experiments: CC MM RBG. Performed the experiments: CC MM. Analyzed the data: CC ARJ NDY BEC RSH MAD SAR SR AL RBG. Contributed reagents/materials/analysis tools: MM EMR. Wrote the paper: CC PWS RBG.Attached Files
Published - Cantacessi2010p10522Plos_Neglect_Trop_D.pdf
Supplemental Material - journal.pntd.0000684.s001.xls
Supplemental Material - journal.pntd.0000684.s002.xls
Supplemental Material - journal.pntd.0000684.s003.xls
Supplemental Material - journal.pntd.0000684.s004.xls
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Additional details
- PMCID
- PMC2867931
- Eprint ID
- 18917
- Resolver ID
- CaltechAUTHORS:20100707-095001420
- National Human Genome Research Institute
- NIH
- Australian Research Council
- Australian Academy of Science
- Australian-American Fulbright Commission
- Created
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2010-07-09Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field