Unexpectedly broad target recognition of the CRISPR-mediated virus defence system in the archaeon Sulfolobus solfataricus

Autoren:Manica, Andrea; Zebec, Ziga; Steinkellner, Julia; Schleper, Christa
Abstrakt:The hyperthermophilic archaeon Sulfolobus solfataricus carries an extensive array of clustered regularly interspaced short palindromic repeats (CRISPR) systems able to mediate DNA degradation of invading genetic elements when complementarity to the small CRISPR-derived (cr)RNAs is given. Studying virus defence in vivo with recombinant viral variants, we demonstrate here that an unexpectedly high number of mutations are tolerated between the CRISPR-derived guide RNAs (crRNAs) and their target sequences (protospacer). Up to 15 mismatches in the crRNA still led to ∼50% of DNA degradation, when these mutations were outside the 'seed' region. More than 15 mutations were necessary to fully abolished interference. Different from other CRISPR systems investigated in vivo, mutations outside the protospacer region indicated no need for a protospacer adjacent motif sequence to confer DNA interference. However, complementarity of only 3 nucleotides between the repeat-derived 5' handle of the crRNA and nucleotides adjacent to the protospacer enabled self-recognition, i.e. protection of the host locus. Our findings show commonalities and differences among the various CRISPR-mediated defence systems and suggest that they should not merely be perceived as a 'first-barrier-defence system' but may be considered to have a broader mechanism that allows host cells to cope with viruses keeping them at reduced levels.
Sprache:Englisch
Publikationsdatum:2013
Journaltitel:Nucleic Acids Research
Volume:41
Nummer:22
Seiten:10509-10517
Peer reviewed:true
Links:
Digital Object Identifier (DOI):http://dx.doi.org/10.1093/nar/gkt767
Publikationstyp:Artikel
Portal:https://ucris.univie.ac.at/portal/en/publications/unexpectedly-broad-target-recognition-of-the-crisprmediated-virus-defence-system-in-the-archaeon-sulfolobus-solfataricus(0531da49-b8ee-4ed4-9740-6864229dee4f).html