Apr 18, 2020

Internal translation of large subunit transcripts drives small subunit synthesis in type I CRISPR-Cas interference complexes

BioRxiv : the Preprint Server for Biology
T. M. McBrideRobert D Fagerlund

Abstract

CRISPR-Cas adaptive immune systems provide bacteria and archaea with defense against their viruses and other mobile genetic elements. CRISPR-Cas immunity involves the formation of ribonucleoprotein complexes that specifically bind and degrade foreign nucleic acids. Despite advances in the biotechnological exploitation of select systems, multiple CRISPR-Cas types remain uncharacterized. Here, we investigated a type I-D system from Synechocystis and revealed the Cascade complex, which is required for interference, forms a hybrid ribonucleoprotein complex that is structurally and genetically related to both type I and III systems. Surprisingly, the type I-D complex contained multiple functionally-important small subunit proteins encoded from an internal in-frame translation initiation site within the large subunit gene, cas10d. Structural analysis revealed that these small subunits bound the complex similarly to Cas11 small subunits in other type I and III systems, where they are encoded as a separate gene. We show that internal translation of small subunits from within large subunit genes is conserved across diverse type I-D, I-B and I-C systems, which account for ~23% of all sequenced CRISPR-Cas systems. Indeed, we demonstrate t...Continue Reading

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