Comparing SSB-PriA Functional and Physical Interactions in Gram-Positive and -Negative Bacteria

Methods Mol Biol. 2021:2281:67-80. doi: 10.1007/978-1-0716-1290-3_4.

Abstract

Single-stranded DNA (ssDNA)-binding protein (SSB) is essential for DNA metabolic processes. SSB also binds to many DNA-binding proteins that constitute the SSB interactome. The mechanism through which PriA helicase, an initiator protein in the DNA replication restart process, is stimulated by SSB in Escherichia coli (EcSSB) has been established. However, some Gram-positive bacterial SSBs such as Bacillus subtilis SsbA (a counterpart of EcSSB), Staphylococcus aureus SsbA, SsbB, and SsbC do not activate PriA helicase. Here, we describe some of the methods used in our laboratory to compare SSB-PriA functional and physical interactions in Gram-positive and -negative bacteria.

Keywords: ATPase; ConSurf; PriA; Protein-protein interaction; SPR; SSB; SsbA; Stimulation; ssDNA-binding protein.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Binding Sites
  • DNA Helicases / chemistry
  • DNA Helicases / metabolism*
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism
  • Models, Molecular
  • Protein Binding
  • Sequence Analysis, Protein
  • Species Specificity
  • Staphylococcus aureus / metabolism*
  • Surface Plasmon Resonance

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • SSB protein, E coli
  • priA protein, E coli
  • DNA Helicases