title: Identification and characterization of a single-stranded DNA-binding protein from the archaeon Methanococcus jannaschii.
abstract: Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria and eukarya. We report here the identification and characterization of the SSB of an archaeon, Methanococcus jannaschii. The M. jannaschii SSB (mjaSSB) has significant amino acid sequence similarity to the eukaryotic SSB, replication protein A (RPA), and contains four tandem repeats of the core single-stranded DNA (ssDNA) binding domain originally defined by structural studies of RPA. Homologous SSBs are encoded by the genomes of other archaeal species, including Methanobacterium thermoautotrophicum and Archaeoglobus fulgidus. The purified mjaSSB binds to ssDNA with high affinity and selectivity. The apparent association constant for binding to ssDNA is similar to that of RPA under comparable experimental conditions, and the affinity for ssDNA exceeds that for double-stranded DNA by at least two orders of magnitude. The binding site size for mjaSSB is approximately 20 nucleotides. Given that RPA is related to mjaSSB at the sequence level and to Escherichia coli SSB at the structural level, we conclude that the SSBs of archaea, eukarya, and bacteria share a common core ssDNA-binding domain. This ssDNA-binding domain was presumably present in the common ancestor to all three major branches of life.
authors.affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21210, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: Kelly
authors.forename: P
authors.initials: P
authors.lastname: Simancek
authors.forename: G S
authors.initials: GS
authors.lastname: Brush
mesh_terms: D000595:Amino Acid Sequence; D019641:DNA, Archaeal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017017:Methanococcus / Q000378:metabolism*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017421:Sequence Analysis
chemical_list: D019641:DNA, Archaeal; D004268:DNA-Binding Proteins