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TitleTarget preference of Type III-A CRISPR-Cas complexes at the transcription bubble.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 3001-3001, Year 2019
Publish dateJul 5, 2019
AuthorsTina Y Liu / Jun-Jie Liu / Abhishek J Aditham / Eva Nogales / Jennifer A Doudna /
PubMed AbstractType III-A CRISPR-Cas systems are prokaryotic RNA-guided adaptive immune systems that use a protein-RNA complex, Csm, for transcription-dependent immunity against foreign DNA. Csm can cleave RNA and ...Type III-A CRISPR-Cas systems are prokaryotic RNA-guided adaptive immune systems that use a protein-RNA complex, Csm, for transcription-dependent immunity against foreign DNA. Csm can cleave RNA and single-stranded DNA (ssDNA), but whether it targets one or both nucleic acids during transcription elongation is unknown. Here, we show that binding of a Thermus thermophilus (T. thermophilus) Csm (TthCsm) to a nascent transcript in a transcription elongation complex (TEC) promotes tethering but not direct contact of TthCsm with RNA polymerase (RNAP). Biochemical experiments show that both TthCsm and Staphylococcus epidermidis (S. epidermidis) Csm (SepCsm) cleave RNA transcripts, but not ssDNA, at the transcription bubble. Taken together, these results suggest that Type III systems primarily target transcripts, instead of unwound ssDNA in TECs, for immunity against double-stranded DNA (dsDNA) phages and plasmids. This reveals similarities between Csm and eukaryotic RNA interference, which also uses RNA-guided RNA targeting to silence actively transcribed genes.
External linksPubMed:31278272 / Publisher's page
KeywordsRNA BINDING PROTEIN/RNA / Csm / RNA substrate / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA complex
MethodsEM (single particle)
Resolution3.8 - 17 A
Structure data

EMDB-0454:
Cryo-EM structure of the T. thermophilus Csm complex bound to target ssRNA

EMDB-0455:
Negative stain EM structure of T. thermophilus Csm with a transcription elongation complex

PDB-6o1o:
Cryo-EM structure of the T. thermophilus Csm complex bound to target ssRNA

Source
  • thermus thermophilus (bacteria)

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