1887

Abstract

Obtaining meaningful viral information from large sequencing datasets presents unique challenges distinct from prokaryotic and eukaryotic sequencing efforts. The difficulties surrounding this issue can be ascribed in part to the genomic plasticity of viruses themselves as well as the scarcity of existing information in genomic databases. The open-source software PhagePhisher (http://www.putonti-lab.com/phagephisher) has been designed as a simple pipeline to extract relevant information from complex and mixed datasets, and will improve the examination of bacteriophages, viruses, and virally related sequences, in a range of environments. Key aspects of the software include speed and ease of use; PhagePhisher can be used with limited operator knowledge of bioinformatics on a standard workstation. As a proof-of-concept, PhagePhisher was successfully implemented with bacteria–virus mixed samples of varying complexity. Furthermore, viral signals within microbial metagenomic datasets were easily and quickly identified by PhagePhisher, including those from prophages as well as lysogenic phages, an important and often neglected aspect of examining phage populations in the environment. PhagePhisher resolves viral-related sequences which may be obscured by or imbedded in bacterial genomes.

Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.000053
2016-03-10
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/mgen/2/3/mgen000053.html?itemId=/content/journal/mgen/10.1099/mgen.0.000053&mimeType=html&fmt=ahah

References

  1. Abeles S. R., Pride D. T. 2014; Molecular bases and role of viruses in the human microbiome. J Mol Biol 426:3892–3906 [View Article][PubMed]
    [Google Scholar]
  2. Akhter S., Aziz R. K., Edwards R. A. 2012; PhiSpy: a novel algorithm for finding prophages in bacterial genomes that combines similarity- and composition-based strategies. Nucleic Acids Res 40:e126 [View Article][PubMed]
    [Google Scholar]
  3. Boetzer M., Henkel C. V., Jansen H. J., Butler D., Pirovano W. 2011; Scaffolding pre-assembled contigs using sspace . Bioinformatics 27:578–579 [View Article][PubMed]
    [Google Scholar]
  4. Bolduc B., Shaughnessy D. P., Wolf Y. I., Koonin E. V., Roberto F. F., Young M. 2012; Identification of novel positive-strand RNA viruses by metagenomic analysis of archaea-dominated Yellowstone hot springs. J Virol 86:5562–5573 [View Article][PubMed]
    [Google Scholar]
  5. Born Y., Fieseler L., Marazzi J., Lurz R., Duffy B., Loessner M. J. 2011; Novel virulent and broad-host-range Erwinia amylovora bacteriophages reveal a high degree of mosaicism and a relationship to Enterobacteriaceae phages. Appl Environ Microbiol 77:5945–5954 [View Article][PubMed]
    [Google Scholar]
  6. Bratbak G., Thingstad F., Heldal M. 1994; Viruses and the microbial loop. Microb Ecol 28:209–221 [View Article][PubMed]
    [Google Scholar]
  7. Clokie M. R., Millard A. D., Letarov A. V., Heaphy S. 2011; Phages in nature. Bacteriophage 1:31–45 [View Article][PubMed]
    [Google Scholar]
  8. Dutilh B. E., Cassman N., McNair K., Sanchez S. E., Silva G. G., Boling L., Barr J. J., Speth D. R., Seguritan V., other authors. 2014; A highly abundant bacteriophage discovered in the unknown sequences of human faecal metagenomes. Nat Commun 5:4498 [View Article][PubMed]
    [Google Scholar]
  9. Hurwitz B. L., Sullivan M. B. 2013; The Pacific Ocean virome (POV): a marine viral metagenomic dataset and associated protein clusters for quantitative viral ecology. PLoS One 8:e57355 [View Article][PubMed]
    [Google Scholar]
  10. Jachiet P. A., Colson P., Lopez P., Bapteste E. 2014; Extensive gene remodeling in the viral world: new evidence for nongradual evolution in the mobilome network. Genome Biol Evol 6:2195–2205 [View Article][PubMed]
    [Google Scholar]
  11. Jacquet S., Miki T., Noble R., Peduzzi P., Wilhelm S. 2010; Viruses in aquatic ecosystems: important advancements of the last 20 years and prospects for the future in the field of microbial oceanography and limnology. Adv Oceanogr Limnol 1:97–141 [View Article]
    [Google Scholar]
  12. Langmead B., Salzberg S. L. 2012; Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359 [View Article][PubMed]
    [Google Scholar]
  13. Lima-Mendez G., Van Helden J., Toussaint A., Leplae R. 2008; Prophinder: a computational tool for prophage prediction in prokaryotic genomes. Bioinformatics 24:863–865 [View Article][PubMed]
    [Google Scholar]
  14. Lu S., Le S., Tan Y., Li M., Liu C., Zhang K., Huang J., Chen H., Rao X., other authors. 2014; Unlocking the mystery of the hard-to-sequence phage genome: PaP1 methylome and bacterial immunity. BMC Genomics 15:803 [View Article][PubMed]
    [Google Scholar]
  15. Malki K., Bruder K., Putonti C. 2015a; Survey of microbial populations within Lake Michigan nearshore waters at two Chicago public beaches. Data Brief 5:556–559 [View Article]
    [Google Scholar]
  16. Malki K., Kula A., Bruder K., Sible E., Hatzopoulos T., Steidel S., Watkins S. C., Putonti C. 2015b; Bacteriophages isolated from Lake Michigan demonstrate broad host-range across several bacterial phyla. Virol J 12:164 [View Article][PubMed]
    [Google Scholar]
  17. Millard A., Clokie M. R., Shub D. A., Mann N. H. 2004; Genetic organization of the psbAD region in phages infecting marine Synechococcus strains. Proc Natl Acad Sci U S A 101:11007–11012 [View Article][PubMed]
    [Google Scholar]
  18. Mizuno C. M., Rodriguez-Valera F., Kimes N. E., Ghai R. 2013; Expanding the marine virosphere using metagenomics. PLoS Genet 9:e1003987 [View Article][PubMed]
    [Google Scholar]
  19. Overbeek R., Olson R., Pusch G. D., Olsen G. J., Davis J. J., Disz T., Edwards R. A., Gerdes S., Parrello B., other authors. 2014; The SEED and the rapid annotation of microbial genomes using subsystems technology (RAST). Nucleic Acids Res 42:(D1)D206–D214 [View Article][PubMed]
    [Google Scholar]
  20. Reyes A., Blanton L. V., Cao S., Zhao G., Manary M., Trehan I., Smith M. I., Wang D., Virgin H. W., other authors. 2015; Gut DNA viromes of Malawian twins discordant for severe acute malnutrition. Proc Natl Acad Sci U S A 112:11941–11946 [View Article][PubMed]
    [Google Scholar]
  21. Rodriguez-Valera F., Mizuno C. M., Ghai R. 2014; Tales from a thousand and one phages. Bacteriophage 4:e28265 [View Article][PubMed]
    [Google Scholar]
  22. Rohwer F., Edwards R. 2002; The phage proteomic tree: a genome-based taxonomy for phage. J Bacteriol 184:4529–4535 [View Article][PubMed]
    [Google Scholar]
  23. Roux S., Enault F., Hurwitz B. L., Sullivan M. B. 2015; VirSorter: mining viral signal from microbial genomic data. PeerJ 3:e985 [View Article][PubMed]
    [Google Scholar]
  24. Santos F., Yarza P., Parro V., Briones C., Antón J. 2010; The metavirome of a hypersaline environment. Environ Microbiol 12:2965–2976 [View Article][PubMed]
    [Google Scholar]
  25. Schmieder R., Edwards R. 2011; Fast identification and removal of sequence contamination from genomic and metagenomic datasets. PLoS One 6:e17288 [View Article][PubMed]
    [Google Scholar]
  26. Watkins S. C., Smith J. R., Hayes P. K., Watts J. E. 2014; Characterisation of host growth after infection with a broad-range freshwater cyanopodophage. PLoS One 9:e87339 [View Article][PubMed]
    [Google Scholar]
  27. Wilhelm S. W., Suttle C. A. 1999; Viruses and nutrient cycles in the sea: viruses play critical roles in the structure and function of aquatic food webs. Bioscience 49:781–788 [View Article]
    [Google Scholar]
  28. Yoshida M., Takaki Y., Eitoku M., Nunoura T., Takai K. 2013; Metagenomic analysis of viral communities in (hado)pelagic sediments. PLoS One 8:e57271 [View Article][PubMed]
    [Google Scholar]
  29. Zerbino D. R., Birney E. 2008; Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 18:821–829 [View Article][PubMed]
    [Google Scholar]
  30. Zhou Y., Liang Y., Lynch K. H., Dennis J. J., Wishart D. S. 2011; phast: a fast phage search tool. Nucleic Acids Res 39:(suppl)W347–W352 [View Article][PubMed]
    [Google Scholar]
  31. 1. Malki, K., Kula, A., Bruder, K., Sible, E., Hatzopoulos, T., Steidel, S., Watkins, S. C. & Putonti, C. GenBank KT254130 (2015).
  32. 2. Tatusova, T., Ciufo, S., Fedorov, B., O'Neill, K., Tolstoy, I. RefSeq microbial genomes database (2014).
http://instance.metastore.ingenta.com/content/journal/mgen/10.1099/mgen.0.000053
Loading
/content/journal/mgen/10.1099/mgen.0.000053
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error