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bioRxiv · 10.1101/2021.03.25.437074

Unlimited cooperativity of Betatectivirus SSB, a novel DNA binding protein related to an atypical group of SSBs from protein-primed replicating bacterial viruses

Lechuga, A.·Kazlauskas, D.·Salas, M.·Redrejo Rodriguez, M.
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Abstract

Bam35 and related betatectiviruses are tail-less bacteriophages that prey on members of the Bacillus cereus group. These temperate viruses replicate their linear genome by a protein-primed mechanism. In this work, we have identified and characterized the product of the viral ORF2 as a single-stranded DNA binding protein (hereafter B35SSB). B35SSB binds ssDNA with great preference over dsDNA or RNA in a sequence-independent, highly cooperative manner that results in a non-specific stimulation of DNA replication. We have also identified several aromatic and basic residues, involved in base-stacking and electrostatic interactions, respectively, that are required for effective protein-ssDNA interaction. Although SSBs are essential for DNA replication in all domains of life as well as many viruses, they are very diverse proteins. However, most SSBs share a common structural domain, named OB-fold. Protein-primed viruses could constitute an exception, as no OB-fold DNA binding protein has been reported. Based on databases searches as well as phylogenetic and structural analyses, we showed that B35SSB belongs to a novel and independent group of SSBs. This group contains proteins encoded by protein-primed viral genomes from unrelated viruses, spanning betatectiviruses and {Phi}29 and close podoviruses, and they share a conserved pattern of secondary structure. Sensitive searches and structural predictions indicate that B35SSB contains a conserved domain resembling a divergent OB-fold, which would constitute the first occurrence of an OB-fold-like domain in a protein-primed genome. HighlightsO_LIBam35 ORF 2 product encodes a viral single-stranded DNA binding protein (B35SSB). C_LIO_LIB35SSB binds ssDNA in a highly cooperative manner but with no sequence specificity. C_LIO_LIB35SSB-ssDNA binding is mediated by base-stacking and ionic interactions. C_LIO_LIBam35 and {Phi}29-related SSBs form a novel group of SSBs from protein-primed viruses. C_LIO_LIThe B35-{Phi}29 SSBs group shares a highly divergent OB-fold-like domain. C_LI

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Lechuga, A., Kazlauskas, D., Salas, M., Redrejo Rodriguez, M.. 2021-03-25. Unlimited cooperativity of Betatectivirus SSB, a novel DNA binding protein related to an atypical group of SSBs from protein-primed replicating bacterial viruses. https://doi.org/10.1101/2021.03.25.437074

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