Purification and characterization of the single-stranded DNA binding protein from Streptococcus pneumoniae

S. E. Steffen, F. R. Bryant

Research output: Contribution to journalArticlepeer-review

Abstract

The Escherichia coli single-stranded DNA binding (SSB) protein is a non-sequence-specific DNA binding protein that functions as an accessory factor for the RecA protein-promoted three-strand exchange reaction. An open reading frame encoding a protein similar in size and sequence to the E. coli SSB protein has been identified in the Streptococcus pneumoniae genome. The open reading frame has been cloned, an overexpression system has been developed, and the protein has been purified to greater than 99% homogeneity. The purified protein binds to ssDNA in a manner similar to that of the E. coli SSB protein. The protein also stimulates the S. pneumoniae RecA protein and E. coli RecA protein-promoted strand exchange reactions to an extent similar to that observed with the E. coli SSB protein. These results indicate that the protein is the S. pneumoniae analog of the E. coli SSB protein. The availability of highly-purified S. pneumoniae SSB protein will facilitate the study of the molecular mechanisms of RecA protein-mediated transformational recombination in S. pneumoniae.

Original languageEnglish (US)
Pages (from-to)165-170
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume388
Issue number1
DOIs
StatePublished - Apr 1 2001

Keywords

  • RecA protein
  • Recombination
  • SSB protein
  • Strand exchange
  • Streptococcus pneumoniae
  • Transformation

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Purification and characterization of the single-stranded DNA binding protein from Streptococcus pneumoniae'. Together they form a unique fingerprint.

Cite this