Primed CRISPR DNA uptake in Pyrococcus furiosus.
Adaptation, Physiological
/ immunology
Base Pairing
Base Sequence
CRISPR-Associated Proteins
/ metabolism
CRISPR-Cas Systems
/ genetics
Clustered Regularly Interspaced Short Palindromic Repeats
/ genetics
DNA
/ genetics
DNA Helicases
/ metabolism
Mutation
Nucleic Acid Hybridization
Plasmids
/ genetics
Pyrococcus furiosus
/ genetics
RNA
/ genetics
Ribonucleoproteins
/ genetics
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
19 06 2020
19 06 2020
Historique:
accepted:
11
05
2020
revised:
15
04
2020
received:
26
01
2020
pubmed:
19
5
2020
medline:
9
9
2020
entrez:
19
5
2020
Statut:
ppublish
Résumé
CRISPR-Cas adaptive immune systems are used by prokaryotes to defend against invaders like viruses and other mobile genetic elements. Immune memories are stored in the form of 'spacers' which are short DNA sequences that are captured from invaders and added to the CRISPR array during a process called 'adaptation'. Spacers are transcribed and the resulting CRISPR (cr)RNAs assemble with different Cas proteins to form effector complexes that recognize matching nucleic acid and destroy it ('interference'). Adaptation can be 'naïve', i.e. independent of any existing spacer matches, or it can be 'primed', i.e. spurred by the crRNA-mediated detection of a complete or partial match to an invader sequence. Here we show that primed adaptation occurs in Pyrococcus furiosus. Although P. furiosus has three distinct CRISPR-Cas interference systems (I-B, I-A and III-B), only the I-B system and Cas3 were necessary for priming. Cas4, which is important for selection and processing of new spacers in naïve adaptation, was also essential for priming. Loss of either the I-B effector proteins or Cas3 reduced naïve adaptation. However, when Cas3 and all crRNP genes were deleted, uptake of correctly processed spacers was observed, indicating that none of these interference proteins are necessary for naïve adaptation.
Identifiants
pubmed: 32421777
pii: 5838857
doi: 10.1093/nar/gkaa381
pmc: PMC7293040
doi:
Substances chimiques
CRISPR-Associated Proteins
0
Ribonucleoproteins
0
RNA
63231-63-0
DNA
9007-49-2
DNA Helicases
EC 3.6.4.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
6120-6135Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM118140
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM118160
Pays : United States
Informations de copyright
© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
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