Target Preference of Type III-A CRISPR-Cas Complexes at the Transcription Bubble

Overview

Journal Nat Commun

Specialty Biology

Date 2019 Jul 7

PMID 31278272

Citations 26

Authors

Tina Y Liu

Jun-Jie Liu

Abhishek J Aditham

Eva Nogales

Jennifer A Doudna

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Mechanistic determinants and dynamics of cA6 synthesis in type III CRISPR-Cas effector complexes.

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Cas10 relieves host growth arrest to facilitate spacer retention during type III-A CRISPR-Cas immunity.

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