The role of Cas8 in type I CRISPR interference
CRISPR (clustered regularly interspaced short palindromic repeat) systems provide bacteria and archaea with adapt- ive immunity to repel invasive genetic elements. Type I systems use ‘cascade’ [CRISPR-associated (Cas) complex for antiviral defence] ribonucleoprotein complexes to target invader DNA,...
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| Format: | Article |
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Portland Press
2015
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| Online Access: | https://eprints.nottingham.ac.uk/35112/ |
| _version_ | 1848795005020274688 |
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| author | Cass, Simon Haas, Karina Stoll, Britta Alkhnbashi, Omer Sharma, Kundan Urlaub, Henning Backofen, Rolf Marchfelder, Anita Bolt, Edward |
| author_facet | Cass, Simon Haas, Karina Stoll, Britta Alkhnbashi, Omer Sharma, Kundan Urlaub, Henning Backofen, Rolf Marchfelder, Anita Bolt, Edward |
| author_sort | Cass, Simon |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | CRISPR (clustered regularly interspaced short palindromic repeat) systems provide bacteria and archaea with adapt- ive immunity to repel invasive genetic elements. Type I systems use ‘cascade’ [CRISPR-associated (Cas) complex for antiviral defence] ribonucleoprotein complexes to target invader DNA, by base pairing CRISPR RNA (crRNA) to protospacers. Cascade identifies PAMs (protospacer adjacent motifs) on invader DNA, triggering R-loop formation and subsequent DNA degradation by Cas3. Cas8 is a candidate PAM recognition factor in some cascades. We analysed Cas8 homologues from type IB CRISPR systems in archaea Haloferax volcanii (Hvo) and Methanothermobacter ther- mautotrophicus (Mth). Cas8 was essential for CRISPR interference in Hvo and purified Mth Cas8 protein responded to PAM sequence when binding to nucleic acids. Cas8 interacted physically with Cas5–Cas7–crRNA complex, stimulating binding to PAM containing substrates. Mutation of conserved Cas8 amino acid residues abolished interference in vivo and altered catalytic activity of Cas8 protein in vitro. This is experimental evidence that Cas8 is important for targeting Cascade to invader DNA. |
| first_indexed | 2025-11-14T19:25:12Z |
| format | Article |
| id | nottingham-35112 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:25:12Z |
| publishDate | 2015 |
| publisher | Portland Press |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-351122020-05-04T17:08:56Z https://eprints.nottingham.ac.uk/35112/ The role of Cas8 in type I CRISPR interference Cass, Simon Haas, Karina Stoll, Britta Alkhnbashi, Omer Sharma, Kundan Urlaub, Henning Backofen, Rolf Marchfelder, Anita Bolt, Edward CRISPR (clustered regularly interspaced short palindromic repeat) systems provide bacteria and archaea with adapt- ive immunity to repel invasive genetic elements. Type I systems use ‘cascade’ [CRISPR-associated (Cas) complex for antiviral defence] ribonucleoprotein complexes to target invader DNA, by base pairing CRISPR RNA (crRNA) to protospacers. Cascade identifies PAMs (protospacer adjacent motifs) on invader DNA, triggering R-loop formation and subsequent DNA degradation by Cas3. Cas8 is a candidate PAM recognition factor in some cascades. We analysed Cas8 homologues from type IB CRISPR systems in archaea Haloferax volcanii (Hvo) and Methanothermobacter ther- mautotrophicus (Mth). Cas8 was essential for CRISPR interference in Hvo and purified Mth Cas8 protein responded to PAM sequence when binding to nucleic acids. Cas8 interacted physically with Cas5–Cas7–crRNA complex, stimulating binding to PAM containing substrates. Mutation of conserved Cas8 amino acid residues abolished interference in vivo and altered catalytic activity of Cas8 protein in vitro. This is experimental evidence that Cas8 is important for targeting Cascade to invader DNA. Portland Press 2015-05-05 Article PeerReviewed Cass, Simon, Haas, Karina, Stoll, Britta, Alkhnbashi, Omer, Sharma, Kundan, Urlaub, Henning, Backofen, Rolf, Marchfelder, Anita and Bolt, Edward (2015) The role of Cas8 in type I CRISPR interference. Bioscience Reports, 35 . e00197/1-e00197/12. ISSN 1573-4935 archaea CRISPR-associated (Cas)8 CRISPR-associated complex for antiviral defence (Cascade) clustered regularly interspaced short palindromic repeat (CRISPR) nuclease http://www.bioscirep.org/content/35/3/e00197 doi:10.1042/BSR20150043 doi:10.1042/BSR20150043 |
| spellingShingle | archaea CRISPR-associated (Cas)8 CRISPR-associated complex for antiviral defence (Cascade) clustered regularly interspaced short palindromic repeat (CRISPR) nuclease Cass, Simon Haas, Karina Stoll, Britta Alkhnbashi, Omer Sharma, Kundan Urlaub, Henning Backofen, Rolf Marchfelder, Anita Bolt, Edward The role of Cas8 in type I CRISPR interference |
| title | The role of Cas8 in type I CRISPR interference |
| title_full | The role of Cas8 in type I CRISPR interference |
| title_fullStr | The role of Cas8 in type I CRISPR interference |
| title_full_unstemmed | The role of Cas8 in type I CRISPR interference |
| title_short | The role of Cas8 in type I CRISPR interference |
| title_sort | role of cas8 in type i crispr interference |
| topic | archaea CRISPR-associated (Cas)8 CRISPR-associated complex for antiviral defence (Cascade) clustered regularly interspaced short palindromic repeat (CRISPR) nuclease |
| url | https://eprints.nottingham.ac.uk/35112/ https://eprints.nottingham.ac.uk/35112/ https://eprints.nottingham.ac.uk/35112/ |