DNA helicase activity of the RecD protein from Deinococcus radiodurans.
The bacterium Deinococcus radiodurans is extremely resistant to high levels of DNA-damaging agents, including gamma rays and ultraviolet light that can lead to double-stranded DNA breaks. Surprisingly, the organism does not appear to have a RecBCD enzyme, an enzyme that is critical for double-strand break repair in many other bacteria. The D. radiodurans genome does encode a protein whose closest characterized homologues are RecD subunits of RecBCD enzymes in other bacteria. We have purified this novel D. radiodurans RecD protein and characterized its biochemical activities. The D. radiodurans RecD protein is a DNA helicase that unwinds short (20 base pairs) DNA duplexes with either a 5'-single-stranded tail or a forked end, but not blunt-ended or 3'-tailed duplexes. Duplexes with 10-12 nucleotide (nt) 5'-tails are good unwinding substrates and are bound tightly, while DNA with shorter tails (4-8 nt) are poor unwinding substrates and are bound much less tightly. The RecD protein is much less efficient at unwinding slightly longer substrates (52 or 76 base pairs, with 12 nt 5'-tails). Unwinding of the longer substrates is stimulated somewhat (4-5-fold) by the single-stranded DNA-binding protein from D. radiodurans. These results show that the D. radiodurans RecD protein is a DNA helicase with 5'-3' polarity and low processivity.
Department of Chemistry & Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
Jianlei
J
Wang
Douglas A
DA
Julin
D000255:Adenosine Triphosphate / Q000378:metabolism; D001483:Base Sequence; D004265:DNA Helicases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D034301:Deinococcus / Q000201:enzymology* / Q000235:genetics; D043211:Exodeoxyribonuclease V / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016680:Genome, Bacterial; D006868:Hydrolysis; D008969:Molecular Sequence Data; D015394:Molecular Structure; D013379:Substrate Specificity
D004269:DNA, Bacterial; D000255:Adenosine Triphosphate; D043211:Exodeoxyribonuclease V; D004265:DNA Helicases