| Summary: | <p>Abstract</p> <p>Background</p> <p><it>Deinococcus radiodurans </it>R1 is one of the most radiation-resistant organisms known and is able to repair an unusually large amount of DNA damage without induced mutation. Single-stranded DNA-binding (SSB) protein is an essential protein in all organisms and is involved in DNA replication, recombination and repair. The published genomic sequence from <it>Deinococcus radiodurans </it>includes a putative single-stranded DNA-binding protein gene (<it>ssb; </it>DR0100) requiring a translational frameshift for synthesis of a complete SSB protein. The apparently tripartite gene has inspired considerable speculation in the literature about potentially novel frameshifting or RNA editing mechanisms. Immediately upstream of the <it>ssb </it>gene is another gene (DR0099) given an <it>ssb</it>-like annotation, but left unexplored.</p> <p>Results</p> <p>A segment of the <it>Deinococcus radiodurans </it>strain R1 genome encompassing the <it>ssb </it>gene has been re-sequenced, and two errors involving omitted guanine nucleotides have been documented. The corrected sequence incorporates both of the open reading frames designated DR0099 and DR0100 into one contiguous <it>ssb </it>open reading frame (ORF). The corrected gene requires no translational frameshifts and contains two predicted oligonucleotide/oligosaccharide-binding (OB) folds. The protein has been purified and its sequence is closely related to the <it>Thermus thermophilus </it>and <it>Thermus aquaticus </it>SSB proteins. Like the <it>Thermus </it>SSB proteins, the SSB<sub>Dr </sub>functions as a homodimer. The <it>Deinococcus radiodurans </it>SSB homodimer stimulates <it>Deinococcus radiodurans </it>RecA protein and <it>Escherichia coli </it>RecA protein-promoted DNA three-strand exchange reactions with at least the same efficiency as the <it>Escherichia coli </it>SSB homotetramer.</p> <p>Conclusions</p> <p>The correct <it>Deinococcus radiodurans ssb </it>gene is a contiguous open reading frame that codes for the largest bacterial SSB monomer identified to date. The <it>Deinococcus radiodurans </it>SSB protein includes two OB folds per monomer and functions as a homodimer. The <it>Deinococcus radiodurans </it>SSB protein efficiently stimulates <it>Deinococcus radiodurans </it>RecA and also <it>Escherichia coli </it>RecA protein-promoted DNA strand exchange reactions. The identification and purification of <it>Deinococcus radiodurans </it>SSB protein not only allows for greater understanding of the SSB protein family but provides an essential yet previously missing player in the current efforts to understand the extraordinary DNA repair capacity of <it>Deinococcus radiodurans</it>.</p>
|
|---|
