SMV1 virus-induced CRISPR spacer acquisition from the conjugative plasmid pMGB1 in Sulfolobus solfataricus P2.

Content Provider	Europe PMC
Author	Erdmann, Susanne Shah, Shiraz A. Garrett, Roger A.
Copyright Year	2013
Abstract	Organisms of the crenarchaeal order Sulfolobales carry complex CRISPR (clustered regularly interspaced short palindromic repeats) adaptive immune systems. These systems are modular and show extensive structural and functional diversity, especially in their interference complexes. The primary targets are an exceptional range of diverse viruses, many of which propagate stably within cells and follow lytic life cycles without producing cell lysis. These properties are consistent with the difficulty of activating CRISPR spacer uptake in the laboratory, but appear to conflict with the high complexity and diversity of the CRISPR immune systems that are found among the Sulfolobales. In the present article, we re-examine the first successful induction of archaeal spacer acquisition in our laboratory that occurred exclusively for the conjugative plasmid pMGB1 in Sulfolobus solfataricus P2 that was co-infected with the virus SMV1 (Sulfolobus monocaudavirus 1). Although we reaffirm that protospacer selection is essentially a random process with respect to the pMGB1 genome, we identified single spacer sequences specific for each of CRISPR loci C, D and E that, exceptionally, occurred in many sequenced clones. Moreover, the same sequence was reproducibly acquired for a given locus in independent experiments, consistent with it being the first protospacer to be selected. There was also a small protospacer bias (1.6:1) to the antisense strand of protein genes. In addition, new experiments demonstrated that spacer acquisition in the previously inactive CRISPR locus A could be induced on freeze–thawing of the infected cells, suggesting that environmental stress can facilitate activation. Coincidentally with spacer acquisition, a mobile OrfB element was deleted from pMGB1, suggesting that interplay can occur between spacer acquisition and transposition.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC3839810&blobtype=pdf
Page Count	10
ISSN	03005127
Journal	Biochemical Society Transactions [Biochem Soc Trans]
Volume Number	41
PubMed Central reference number	PMC3839810
Issue Number	Pt 6
Issue Number	6
PubMed reference number	24256236
e-ISSN	14708752
DOI	10.1042/bst20130196
Language	English
Publisher	Portland Press Ltd.
Publisher Date	2013-12-01
Access Restriction	Open
Rights License	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 work is properly cited. © 2013 The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Licence (CC-BY)(http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
Subject Keyword	Archaea clustered regularly interspaced short palindromic repeats spacer (CRISPR spacer) pMGB1 Sulfolobus Sulfolobus monocaudavirus 1 (SMV1) transposable element ATV, Acidianus two-tailed virus Cas, CRISPR-associated CRISPR, clustered regularly interspaced short palindromic repeats crRNA, CRISPR RNA IS, insertion sequence PAM, protospacer-adjacent motif p.i., post-infection SIRV, Sulfolobus islandicus rod-shaped virus SMV1, Sulfolobus monocaudavirus 1 SSV, Sulfolobus spindle-shaped virus STIV, Sulfolobus turreted icosahedral virus STSV1, Sulfolobus tengchongensis spindle-shaped virus 1
Content Type	Text
Resource Type	Article
Subject	Biochemistry