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Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector‐borne viral and plasmid genes and protospacers
Summary The adaptive immune CRISPR/Cas and CRISPR/Cmr systems of the crenarchaeal thermoacidophile Sulfolobus were challenged by a variety of viral and plasmid genes, and protospacers preceded by different dinucleotide motifs. The genes and protospacers were constructed to carry sequences matching i...
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| Published in: | Molecular microbiology 2011-01, Vol.79 (1), p.35-49 |
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| container_title | Molecular microbiology |
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| creator | Gudbergsdottir, Soley Deng, Ling Chen, Zhengjun Jensen, Jaide V. K. Jensen, Linda R. She, Qunxin Garrett, Roger A. |
| description | Summary
The adaptive immune CRISPR/Cas and CRISPR/Cmr systems of the crenarchaeal thermoacidophile Sulfolobus were challenged by a variety of viral and plasmid genes, and protospacers preceded by different dinucleotide motifs. The genes and protospacers were constructed to carry sequences matching individual spacers of CRISPR loci, and a range of mismatches were introduced. Constructs were cloned into vectors carrying pyrE/pyrF genes and transformed into uracil auxotrophic hosts derived from Sulfolobus solfataricus P2 or Sulfolobus islandicus REY15A. Most constructs, including those carrying different protospacer mismatches, yielded few viable transformants. These were shown to carry either partial deletions of CRISPR loci, covering a broad spectrum of sizes and including the matching spacer, or deletions of whole CRISPR/Cas modules. The deletions occurred independently of whether genes or protospacers were transcribed. For family I CRISPR loci, the presence of the protospacer CC motif was shown to be important for the occurrence of deletions. The results are consistent with a low level of random dynamic recombination occurring spontaneously, either inter‐genomically or intra‐genomically, at the repeat regions of Sulfolobus CRISPR loci. Moreover, the relatively high incidence of single‐spacer deletions observed for S. islandicus suggests that an additional more directed mechanism operates in this organism. |
| doi_str_mv | 10.1111/j.1365-2958.2010.07452.x |
| format | article |
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The adaptive immune CRISPR/Cas and CRISPR/Cmr systems of the crenarchaeal thermoacidophile Sulfolobus were challenged by a variety of viral and plasmid genes, and protospacers preceded by different dinucleotide motifs. The genes and protospacers were constructed to carry sequences matching individual spacers of CRISPR loci, and a range of mismatches were introduced. Constructs were cloned into vectors carrying pyrE/pyrF genes and transformed into uracil auxotrophic hosts derived from Sulfolobus solfataricus P2 or Sulfolobus islandicus REY15A. Most constructs, including those carrying different protospacer mismatches, yielded few viable transformants. These were shown to carry either partial deletions of CRISPR loci, covering a broad spectrum of sizes and including the matching spacer, or deletions of whole CRISPR/Cas modules. The deletions occurred independently of whether genes or protospacers were transcribed. For family I CRISPR loci, the presence of the protospacer CC motif was shown to be important for the occurrence of deletions. The results are consistent with a low level of random dynamic recombination occurring spontaneously, either inter‐genomically or intra‐genomically, at the repeat regions of Sulfolobus CRISPR loci. Moreover, the relatively high incidence of single‐spacer deletions observed for S. islandicus suggests that an additional more directed mechanism operates in this organism.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1111/j.1365-2958.2010.07452.x</identifier><identifier>PMID: 21166892</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Bacteria ; DNA, Bacterial - genetics ; DNA, Bacterial - metabolism ; DNA, Intergenic - genetics ; DNA, Intergenic - metabolism ; DNA, Viral - genetics ; DNA, Viral - metabolism ; Genes ; Genes, Bacterial ; Genes, Viral ; Genomics ; Microbial Viability ; Microbiology ; Organismal biology ; Plasmids ; Recombination, Genetic ; Repetitive Sequences, Nucleic Acid ; Sequence Deletion ; Sulfolobus ; Sulfolobus solfataricus - genetics ; Sulfolobus solfataricus - metabolism ; Transformation, Genetic ; Viruses</subject><ispartof>Molecular microbiology, 2011-01, Vol.79 (1), p.35-49</ispartof><rights>2010 Blackwell Publishing Ltd</rights><rights>2010 Blackwell Publishing Ltd.</rights><rights>Copyright Blackwell Publishing Ltd. Jan 2011</rights><rights>Copyright © 2011 Blackwell Publishing Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5322-185221cfdbbc5322239e48b01ab12db5db38fb957d234846ffebf19b4fd647903</citedby><cites>FETCH-LOGICAL-c5322-185221cfdbbc5322239e48b01ab12db5db38fb957d234846ffebf19b4fd647903</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-2958.2010.07452.x$$EPDF$$P50$$Gwiley$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-2958.2010.07452.x$$EHTML$$P50$$Gwiley$$Hfree_for_read</linktohtml><link.rule.ids>230,315,786,790,891,27957,27958,50923,51032</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21166892$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gudbergsdottir, Soley</creatorcontrib><creatorcontrib>Deng, Ling</creatorcontrib><creatorcontrib>Chen, Zhengjun</creatorcontrib><creatorcontrib>Jensen, Jaide V. K.</creatorcontrib><creatorcontrib>Jensen, Linda R.</creatorcontrib><creatorcontrib>She, Qunxin</creatorcontrib><creatorcontrib>Garrett, Roger A.</creatorcontrib><title>Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector‐borne viral and plasmid genes and protospacers</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
The adaptive immune CRISPR/Cas and CRISPR/Cmr systems of the crenarchaeal thermoacidophile Sulfolobus were challenged by a variety of viral and plasmid genes, and protospacers preceded by different dinucleotide motifs. The genes and protospacers were constructed to carry sequences matching individual spacers of CRISPR loci, and a range of mismatches were introduced. Constructs were cloned into vectors carrying pyrE/pyrF genes and transformed into uracil auxotrophic hosts derived from Sulfolobus solfataricus P2 or Sulfolobus islandicus REY15A. Most constructs, including those carrying different protospacer mismatches, yielded few viable transformants. These were shown to carry either partial deletions of CRISPR loci, covering a broad spectrum of sizes and including the matching spacer, or deletions of whole CRISPR/Cas modules. The deletions occurred independently of whether genes or protospacers were transcribed. For family I CRISPR loci, the presence of the protospacer CC motif was shown to be important for the occurrence of deletions. The results are consistent with a low level of random dynamic recombination occurring spontaneously, either inter‐genomically or intra‐genomically, at the repeat regions of Sulfolobus CRISPR loci. Moreover, the relatively high incidence of single‐spacer deletions observed for S. islandicus suggests that an additional more directed mechanism operates in this organism.</description><subject>Bacteria</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA, Intergenic - genetics</subject><subject>DNA, Intergenic - metabolism</subject><subject>DNA, Viral - genetics</subject><subject>DNA, Viral - metabolism</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genes, Viral</subject><subject>Genomics</subject><subject>Microbial Viability</subject><subject>Microbiology</subject><subject>Organismal biology</subject><subject>Plasmids</subject><subject>Recombination, Genetic</subject><subject>Repetitive Sequences, Nucleic Acid</subject><subject>Sequence Deletion</subject><subject>Sulfolobus</subject><subject>Sulfolobus solfataricus - genetics</subject><subject>Sulfolobus solfataricus - metabolism</subject><subject>Transformation, Genetic</subject><subject>Viruses</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><recordid>eNqNkc2O0zAQxyMEYsvCKyCLC6d0_REnzgEkVL4q7Qq0CxI3y07GrSsnDnbSbm88AhJvyJOQtEsFnPDF9sx_fjOjf5IggudkPBebOWE5T2nJxZziMYqLjNP57b1kdkrcT2a45Dhlgn45Sx7FuMGYMJyzh8kZJSTPRUlnyY_X-1Y1tkJd8B2E3kJE3qB-DehmcMY7r4eIFtfLm4_XFwsVkWrr07cJKO5jD01EuzW0qFor56BdQY12tl-jLVS9Dz-_fdc-tIC2Nih3AHROxcbWaAUtHJFj-97HTlUQ4uPkgVEuwpO7-zz5_PbNp8X79PLDu-Xi1WVacUZpSgSnlFSm1voQoKyETGhMlCa01rzWTBhd8qKmLBNZbgxoQ0qdmTrPihKz8-TlkdsNuoG6grYfB5RdsI0Ke-mVlX9nWruWK7-VDFNOiBgBz-8AwX8dIPaysbEC51QLfohS8LwockqmVs_-UW78ENpxOykyzATjeBKJo6gKPsYA5jQKwXLyXW7kZK-c7JWT7_Lgu7wdS5_-ucqp8LfRo-DFUbCzDvb_DZZXV8vpxX4BKfW_ow</recordid><startdate>201101</startdate><enddate>201101</enddate><creator>Gudbergsdottir, Soley</creator><creator>Deng, Ling</creator><creator>Chen, Zhengjun</creator><creator>Jensen, Jaide V. K.</creator><creator>Jensen, Linda R.</creator><creator>She, Qunxin</creator><creator>Garrett, Roger A.</creator><general>Blackwell Publishing Ltd</general><scope>24P</scope><scope>WIN</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>201101</creationdate><title>Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector‐borne viral and plasmid genes and protospacers</title><author>Gudbergsdottir, Soley ; Deng, Ling ; Chen, Zhengjun ; Jensen, Jaide V. K. ; Jensen, Linda R. ; She, Qunxin ; Garrett, Roger A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5322-185221cfdbbc5322239e48b01ab12db5db38fb957d234846ffebf19b4fd647903</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Bacteria</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - metabolism</topic><topic>DNA, Intergenic - genetics</topic><topic>DNA, Intergenic - metabolism</topic><topic>DNA, Viral - genetics</topic><topic>DNA, Viral - metabolism</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>Genes, Viral</topic><topic>Genomics</topic><topic>Microbial Viability</topic><topic>Microbiology</topic><topic>Organismal biology</topic><topic>Plasmids</topic><topic>Recombination, Genetic</topic><topic>Repetitive Sequences, Nucleic Acid</topic><topic>Sequence Deletion</topic><topic>Sulfolobus</topic><topic>Sulfolobus solfataricus - genetics</topic><topic>Sulfolobus solfataricus - metabolism</topic><topic>Transformation, Genetic</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gudbergsdottir, Soley</creatorcontrib><creatorcontrib>Deng, Ling</creatorcontrib><creatorcontrib>Chen, Zhengjun</creatorcontrib><creatorcontrib>Jensen, Jaide V. K.</creatorcontrib><creatorcontrib>Jensen, Linda R.</creatorcontrib><creatorcontrib>She, Qunxin</creatorcontrib><creatorcontrib>Garrett, Roger A.</creatorcontrib><collection>Wiley Online Library Open Access</collection><collection>Wiley Online Library Free Content</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gudbergsdottir, Soley</au><au>Deng, Ling</au><au>Chen, Zhengjun</au><au>Jensen, Jaide V. K.</au><au>Jensen, Linda R.</au><au>She, Qunxin</au><au>Garrett, Roger A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector‐borne viral and plasmid genes and protospacers</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>2011-01</date><risdate>2011</risdate><volume>79</volume><issue>1</issue><spage>35</spage><epage>49</epage><pages>35-49</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><notes>Present address: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.</notes><notes>Re‐use of this article is permitted in accordance with the Terms and Conditions set out at</notes><notes>These authors contributed equally to this article.</notes><notes>http://wileyonlinelibrary.com/onlineopen#OnlineOpen_Terms</notes><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><notes>These authors contributed equally to this article.Re-use of this article is permitted in accordance with the Terms and Conditions set out at http://wileyonlinelibrary.com/onlineopen#OnlineOpen_Terms</notes><abstract>Summary
The adaptive immune CRISPR/Cas and CRISPR/Cmr systems of the crenarchaeal thermoacidophile Sulfolobus were challenged by a variety of viral and plasmid genes, and protospacers preceded by different dinucleotide motifs. The genes and protospacers were constructed to carry sequences matching individual spacers of CRISPR loci, and a range of mismatches were introduced. Constructs were cloned into vectors carrying pyrE/pyrF genes and transformed into uracil auxotrophic hosts derived from Sulfolobus solfataricus P2 or Sulfolobus islandicus REY15A. Most constructs, including those carrying different protospacer mismatches, yielded few viable transformants. These were shown to carry either partial deletions of CRISPR loci, covering a broad spectrum of sizes and including the matching spacer, or deletions of whole CRISPR/Cas modules. The deletions occurred independently of whether genes or protospacers were transcribed. For family I CRISPR loci, the presence of the protospacer CC motif was shown to be important for the occurrence of deletions. The results are consistent with a low level of random dynamic recombination occurring spontaneously, either inter‐genomically or intra‐genomically, at the repeat regions of Sulfolobus CRISPR loci. Moreover, the relatively high incidence of single‐spacer deletions observed for S. islandicus suggests that an additional more directed mechanism operates in this organism.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21166892</pmid><doi>10.1111/j.1365-2958.2010.07452.x</doi><tpages>15</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Molecular microbiology, 2011-01, Vol.79 (1), p.35-49 |
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| subjects | Bacteria DNA, Bacterial - genetics DNA, Bacterial - metabolism DNA, Intergenic - genetics DNA, Intergenic - metabolism DNA, Viral - genetics DNA, Viral - metabolism Genes Genes, Bacterial Genes, Viral Genomics Microbial Viability Microbiology Organismal biology Plasmids Recombination, Genetic Repetitive Sequences, Nucleic Acid Sequence Deletion Sulfolobus Sulfolobus solfataricus - genetics Sulfolobus solfataricus - metabolism Transformation, Genetic Viruses |
| title | Dynamic properties of the Sulfolobus CRISPR/Cas and CRISPR/Cmr systems when challenged with vector‐borne viral and plasmid genes and protospacers |
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