Advertisement

CRISPR pp 223-232 | Cite as

Archaeal Viruses of the Sulfolobales: Isolation, Infection, and CRISPR Spacer Acquisition

  • Susanne ErdmannEmail author
  • Roger A. Garrett
Part of the Methods in Molecular Biology book series (MIMB, volume 1311)

Abstract

Infection of archaea with phylogenetically diverse single viruses, performed in different laboratories, has failed to activate spacer acquisition into host CRISPR loci. The first successful uptake of archaeal de novo spacers was observed on infection of Sulfolobus solfataricus P2 with an environmental virus mixture isolated from Yellowstone National Park (Erdmann and Garrett, Mol Microbiol 85:1044–1056, 2012). Experimental studies of isolated genetic elements from this mixture revealed that SMV1 (S ulfolobus Monocauda Virus 1), a tailed spindle-shaped virus, can induce spacer acquisition in CRISPR loci of Sulfolobus species from a second coinfecting conjugative plasmid or virus (Erdmann and Garrett, Mol Microbiol 85:1044–1056, 2012; Erdmann et al. Mol Microbiol 91:900–917, 2014). Here we describe, firstly, the isolation of archaeal virus mixtures from terrestrial hot springs and the techniques used both to infect laboratory strains with these virus mixtures and to obtain purified virus particles. Secondly, we present the experimental conditions required for activating SMV1-induced spacer acquisition in two different Sulfolobus species.

Key words

CRISPR Spacer acquisition Archaeal Virus Conjugative plasmid Sulfolobales 

Notes

Acknowledgements

The work was supported by grants from the Danish Natural Science Research Council and Copenhagen University. We are grateful to Dr. Xu Peng and Soley Ruth Gudbergsdottir for helpful advice and discussions.

References

  1. 1.
    Prangishvili D, Forterre P, Garrett RA (2006) Viruses of the Archaea: a unifying view. Nat Rev Microbiol 4:837–848CrossRefPubMedGoogle Scholar
  2. 2.
    Gudbergsdottir S, Deng L, Chen Z, Jensen JVK, Jensen LR, She Q et al (2011) Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector-borne viral and plasmid genes and protospacers. Mol Microbiol 79:35–49CrossRefPubMedCentralPubMedGoogle Scholar
  3. 3.
    Manica A, Zebec Z, Teichmann D, Schleper C (2011) In vivo activity of CRISPR-mediated virus defence in a hyperthermophilic archaeon. Mol Microbiol 80:481–491CrossRefPubMedGoogle Scholar
  4. 4.
    Zhang J, Rouillon C, Kerou M, Reeks J, Brügger K, Graham SJ et al (2012) Structure and mechanism of the CMR complex for CRISPR-mediated antiviral immunity. Mol Cell 45:303–313CrossRefPubMedCentralPubMedGoogle Scholar
  5. 5.
    Deng L, Garrett RA, Shah SA, Peng X, She Q (2013) A novel interference mechanism by a type IIIB CRISPR-Cmr module in Sulfolobus. Mol Microbiol 87:1088–1099CrossRefPubMedGoogle Scholar
  6. 6.
    Garrett RA, Vestergaard G, Shah SA (2011) Archaeal CRISPR-based immune systems: exchangeable functional modules. Trends Microbiol 19:549–556CrossRefPubMedGoogle Scholar
  7. 7.
    Erdmann S, Garrett RA (2012) Selective and hyperactive uptake of foreign DNA by adaptive immune systems of an archaeon via two distinct mechanisms. Mol Microbiol 86:757, Mol Microbiol 85: 1044–1056. CorrigendumCrossRefGoogle Scholar
  8. 8.
    Erdmann S, Le Moine Bauer S, Garrett RA (2014) Inter-viral conflicts that exploit host CRISPR immune systems of Sulfolobus. Mol Microbiol 91:900–917CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Archaea Centre, Department of BiologyUniversity of CopenhagenCopenhagen NDenmark
  2. 2.School of Biotechnology and Biomolecular SciencesUniversity of New South WalesSydneyAustralia

Personalised recommendations