Structure–function relationships of two paralogous single-stranded DNA-binding proteins from Streptomyces coelicolor : implication of SsbB in chromosome segregation during sporulation

Paradzik, Tina and Ivic, Nives and Filic, Zelimira and Manjasetty, Babu and Herron, Paul and Luic, Marija and Vujaklija, Dusica (2013) Structure–function relationships of two paralogous single-stranded DNA-binding proteins from Streptomyces coelicolor : implication of SsbB in chromosome segregation during sporulation. Nucleic Acids Research, 41 (6). pp. 3659-3672. ISSN 0305-1048

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Abstract

The linear chromosome of Streptomyces coelicolor contains two paralogous ssb genes, ssbA and ssbB. Following mutational analysis, we concluded that ssbA is essential, whereas ssbB plays a key role in chromosome segregation during sporulation. In the ssbB mutant, 0% of spores lacked DNA. The two ssb genes were expressed differently; in minimal medium, gene expression was prolonged for both genes and significantly upregulated for ssbB. The ssbA gene is transcribed as part of a polycistronic mRNA from two initiation sites, 163 bp and 75 bp upstream of the rpsF translational start codon. The sbB gene is transcribed as a monocistronic mRNA, from an unusual promoter region, 73 bp upstream of the AUG codon. Distinctive DNA-binding affinities of single-stranded DNA-binding proteins monitored by tryptophan fluorescent quenching and electrophoretic mobility shift were observed. The crystal structure of SsbB at 1.7A ° resolution revealed a common OB-fold, lack of the clamp-like structure conserved in SsbA and previously unpublished S-S bridges between the A/B and C/D subunits. This is the first report of the determination of paralogous singlestranded DNA-binding protein structures from the same organism. Phylogenetic analysis revealed frequent duplication of ssb genes in Actinobacteria, whereas their strong retention suggests that they are involved in important cellular functions.