Replication Protein Rep Provides Selective Advantage to Viruses in the Presence of CRISPR-Cas Immunity

CRISPR J. 2023 Feb;6(1):32-42. doi: 10.1089/crispr.2022.0037. Epub 2022 Dec 22.

Abstract

Anti-Clustered regularly interspaced small palindromic repeat (CRISPR) (Acr) phages cooperate to establish a successful infection in CRISPR-containing host. We report here the selective advantage provided by a replication initiator, Rep, toward cooperative host immunosuppression by viruses encoding Acrs. A rep knockout mutant (Δgp16) of Sulfolobus islandicus rod-shaped virus 2 produced around fourfold less virus in a CRISPR-null host, suggesting that Rep is the major replication initiator. In addition to Rep-dependent replication initiation from the viral genomic termini, we detected Rep-independent replication initiation from nonterminal sites. Intriguingly, despite the presence of Acrs, lack of Rep showed a profound effect on virus propagation in a host carrying CRISPR-Cas immunity. Accordingly, the co-infecting parental virus (rep-containing) outcompeted the Δgp16 mutant much more quickly in the CRISPR-containing host than in CRISPR-null host. Despite the nonessentiality, rep is carried by all known members of Rudiviridae, which is likely an evolutionary outcome driven by the ubiquitous presence of CRISPR-Cas in Sulfolobales.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophages* / genetics
  • CRISPR-Cas Systems / genetics
  • Gene Editing
  • Sulfolobus* / genetics
  • Viruses* / genetics