The glycine-rich flexible region in SSB is crucial for PriA stimulation. Issue 61 (15th October 2018)

• Record Type: Journal Article
• Authors: Huang, Yen-Hua; Huang, Cheng-Yang
• Is Part Of: RSC advances. Volume 8:Issue 61(2018)
• Page Start: 35280
• Page End: 35288
• Publication Date: 2018-10-15
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c8ra07306f
• Languages: English
• ISSNs: 2046-2069

Abstract

Abstract : Single-stranded DNA-binding protein (SSB) is essential for all DNA-dependent cellular processes. Abstract : Single-stranded DNA-binding protein (SSB) is essential for all DNA-dependent cellular processes. The mechanism through which PriA helicase, an initiator protein in the DNA replication restart process, is stimulated by SSB in Escherichia coli (Ec) has been established. Nevertheless, whether or not PriA stimulated by SSB is conserved among Gram-negative bacteria remains unclear, and the SSB specificity for the stimulation effect on PriA is unknown. In this study, three similar SSBs from Klebsiella pneumoniae (KpSSB), Salmonella enterica (StSSB), and Pseudomonas aeruginosa (PaSSB) were used to analyze the stimulation effect. Two chimeric proteins, namely, KpSSBn-PaSSBc and KpSSBn-StSSBc, were also used. KpSSB, StSSB, and KpSSBn-StSSBc can stimulate KpPriA activity, but PaSSB and KpSSBn-PaSSBc cannot. The crystal structure of PaSSB solved at 2.04 Å resolution (PDB entry ;5YUO ) reveals the classic OB fold structure, similar to that of EcSSB. Comparison of SSBs through sequence analysis showed that the typical glycine-rich flexible region in PaSSB contains very few glycine residues. Through analyses of protein chimeragenesis, structure–sequence, and ATPase stimulation effects, we concluded that the inherent difference in the glycine-rich flexible region among SSB species is a determinant of PriA stimulation. Further research can directly focus on determining the type of glycine-rich hinge that can stimulate PriA and the reason why bacterial SSBs need to evolve different C-terminal domains during evolution.