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TitleMultiple adaptations underly co-option of a CRISPR surveillance complex for RNA-guided DNA transposition.
Journal, issue, pagesMol Cell, Vol. 83, Issue 11, Page 1827-11838.e6, Year 2023
Publish dateJun 1, 2023
AuthorsJung-Un Park / Michael T Petassi / Shan-Chi Hsieh / Eshan Mehrotra / Gabriel Schuler / Jagat Budhathoki / Vinh H Truong / Summer B Thyme / Ailong Ke / Elizabeth H Kellogg / Joseph E Peters /
PubMed AbstractCRISPR-associated transposons (CASTs) are natural RNA-directed transposition systems. We demonstrate that transposon protein TniQ plays a central role in promoting R-loop formation by RNA-guided DNA- ...CRISPR-associated transposons (CASTs) are natural RNA-directed transposition systems. We demonstrate that transposon protein TniQ plays a central role in promoting R-loop formation by RNA-guided DNA-targeting modules. TniQ residues, proximal to CRISPR RNA (crRNA), are required for recognizing different crRNA categories, revealing an unappreciated role of TniQ to direct transposition into different classes of crRNA targets. To investigate adaptations allowing CAST elements to utilize attachment sites inaccessible to CRISPR-Cas surveillance complexes, we compared and contrasted PAM sequence requirements in both I-F3b CAST and I-F1 CRISPR-Cas systems. We identify specific amino acids that enable a wider range of PAM sequences to be accommodated in I-F3b CAST elements compared with I-F1 CRISPR-Cas, enabling CAST elements to access attachment sites as sequences drift and evade host surveillance. Together, this evidence points to the central role of TniQ in facilitating the acquisition of CRISPR effector complexes for RNA-guided DNA transposition.
External linksMol Cell / PubMed:37267904 / PubMed Central
MethodsEM (single particle)
Resolution3.52 - 4.03 Å
Structure data

26348

7u5d

EMDB-26348, PDB-7u5d:
I-F3b Cascade-TniQ full R-loop complex
Method: EM (single particle) / Resolution: 3.52 Å

26349

7u5e

EMDB-26349: I-F3b Cascade-TniQ full R-loop complex
PDB-7u5e: I-F3b Cascade-TniQ partial R-loop complex
Method: EM (single particle) / Resolution: 4.03 Å

Source
  • aeromonas salmonicida (bacteria)
KeywordsDNA BINDING PROTEIN/DNA/RNA / CRISPR-Cas / Transposon / CAST / Cascade / I-F / I-F3 / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA-RNA complex

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