Fig. 6: The SSB-CTT is essential to recruit RecO following replisome inactivation at a single-strand discontinuity.
From: Rescuing the bacterial replisome at a nick requires recombinational repair and helicase reloading
A Schematic of SSB primary structure with corresponding functions. Numbers indicate amino acid residue boundaries between the N-terminal domain (NTD), C-terminal domain (CTD) and C-terminal tail (CTT). B Structural model of B. subtilis SSB homotetramer in the presence of single-strand DNA (AlphaFold 3). A sixty nucleotide homopolymer of thymidine bases (dT60) is shown in black and SSB-CTT residues are highlighted in red. C Spot-titre analysis of SSB variants in strains engineered to express (+ xylose) nCas9. Top panel shows the genetic system employed in this experiment. sgRNAs were targeted to a locus at -90°. D MFA using whole genome sequencing in strains with SSB variants engineered to express nCas9. The frequency of sequencing reads was plotted against genome position. E ChIP-qPCR analyses of SSB, His-RecO and His-RecA proteins in strains engineered to express nCas9. qPCR primers amplified either a site located -3 kb from the sgRNA target (-90°) or a control locus (+90°) used for normalisation. Error bars indicate the standard error of the mean and circles overlaid on bars correspond to three biological replicates. F ChIP-qPCR analyses of His-RecA in strains with recombinational repair mutants engineered to express nCas9. Top panel shows the genetic system employed in this experiment. qPCR primers amplified either a site located -3 kb from the sgRNA target (-90°) or a control locus (+90°) used for normalisation. Error bars indicate the standard error of the mean and circles overlaid on bars correspond to three biological replicates. Source data are provided as a Source Data file.
