Direct CRISPR spacer acquisition from RNA by a natural reverse transcriptase-Cas1 fusion protein

通过天然逆转录酶-Cas1融合蛋白直接从RNA获取CRISPR间隔区

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作者:Sukrit Silas #, Georg Mohr #, David J Sidote, Laura M Markham, Antonio Sanchez-Amat, Devaki Bhaya, Alan M Lambowitz, Andrew Z Fire

Abstract

CRISPR systems mediate adaptive immunity in diverse prokaryotes. CRISPR-associated Cas1 and Cas2 proteins have been shown to enable adaptation to new threats in type I and II CRISPR systems by the acquisition of short segments of DNA (spacers) from invasive elements. In several type III CRISPR systems, Cas1 is naturally fused to a reverse transcriptase (RT). In the marine bacterium Marinomonas mediterranea (MMB-1), we showed that a RT-Cas1 fusion protein enables the acquisition of RNA spacers in vivo in a RT-dependent manner. In vitro, the MMB-1 RT-Cas1 and Cas2 proteins catalyze the ligation of RNA segments into the CRISPR array, which is followed by reverse transcription. These observations outline a host-mediated mechanism for reverse information flow from RNA to DNA.

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