Regulatory sequence-based discovery of anti-defense genes in archaeal viruses

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In silico identification of viral anti-CRISPR proteins (Acrs) has relied largely on the guilt-by-association method using known Acrs or anti-CRISPR associated proteins (Acas) as the bait. However, the low number and limited spread of the characterized archaeal Acrs and Aca hinders our ability to identify Acrs using guilt-by-association. Here, based on the observation that the few characterized archaeal Acrs and Aca are transcribed immediately post viral infection, we hypothesize that these genes, and many other unidentified anti-defense genes (ADG), are under the control of conserved regulatory sequences including a strong promoter, which can be used to predict anti-defense genes in archaeal viruses. Using this consensus sequence based method, we identify 354 potential ADGs in 57 archaeal viruses and 6 metagenome-assembled genomes. Experimental validation identified a CRISPR subtype I-A inhibitor and the first virally encoded inhibitor of an archaeal toxin-antitoxin based immune system. We also identify regulatory proteins potentially akin to Acas that can facilitate further identification of ADGs combined with the guilt-by-association approach. These results demonstrate the potential of regulatory sequence analysis for extensive identification of ADGs in viruses of archaea and bacteria.
OriginalsprogEngelsk
Artikelnummer3699
TidsskriftNature Communications
Vol/bind15
Udgave nummer1
Antal sider13
ISSN2041-1723
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
We thank all members of the Microbial Immunity lab, Department of biology, Copenhagen University for their suggestions. This work was supported by the Novo Nordisk Fonden Postdoctoral Fellowship in Bioscience and Basic Biomedicine Grant (NNF21OC0067491) to Y.B.-C., and by the Danish Council for Independent Research/Natural Sciences [DFF-0135-00402] and Novo Nordisk Foundation/Hallas M\u00F8ller Ascending Investigator Grant [NNF17OC0031154] to X.P.

Publisher Copyright:
© The Author(s) 2024.

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