TY - JOUR
T1 - Cutting of chi-like sequences by the RecBCD enzyme of Escherichia coli
AU - Cheng, Keith C.
AU - Smith, Gerald R.
N1 - Funding Information:
We thank Sue Amundsen, Gary Braedt, Stan Brown, Fred Ponticelli and Andrew Taylor for encouragement and discussion. We are additionally indebted to Andrew Taylor for providing RecBCD enzyme and technical advice. Members of Ron Reeder’s laboratory generously helped with their sequencing facilities and Jim Wallace provided the Genepro DNA sequence restriction program. This research was supported by the NIH through grant GM-31693 to G.R.S. and in part by training grant 5 T32 GM-07187 to K.C.C.
PY - 1987/4/20
Y1 - 1987/4/20
N2 - Chi, 5′ G-C-T-G-G-T-G-G 3′, stimulates coliphage λ recombination mediated by the major recombination pathway of Escherichia coli, the RecBC pathway. Purified RecBCD enzyme makes a single strand endonuclease cleavage four to six nucleotides to the 3′ side of the chi sequence. Three sequences similar to chi, 5′ A-C-T-G-G-T-G-G 3′, 5′ G-T-T-G-G-T-G-G 3′ and 5′ G-C-T-A-G-T-G-G 3′, have partial recombinational hotspot activity in genetic crosses. We report here that purified RecBCD enzyme preferentially cuts four nucleotides to the right of the two most active chi-like octamers. The degree of cutting correlated with the genetic activities of these sequences; this result indicates that these cleavages are essential to the genetic activity of chi and chi-like sequences.
AB - Chi, 5′ G-C-T-G-G-T-G-G 3′, stimulates coliphage λ recombination mediated by the major recombination pathway of Escherichia coli, the RecBC pathway. Purified RecBCD enzyme makes a single strand endonuclease cleavage four to six nucleotides to the 3′ side of the chi sequence. Three sequences similar to chi, 5′ A-C-T-G-G-T-G-G 3′, 5′ G-T-T-G-G-T-G-G 3′ and 5′ G-C-T-A-G-T-G-G 3′, have partial recombinational hotspot activity in genetic crosses. We report here that purified RecBCD enzyme preferentially cuts four nucleotides to the right of the two most active chi-like octamers. The degree of cutting correlated with the genetic activities of these sequences; this result indicates that these cleavages are essential to the genetic activity of chi and chi-like sequences.
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U2 - 10.1016/0022-2836(87)90252-X
DO - 10.1016/0022-2836(87)90252-X
M3 - Article
C2 - 2958631
AN - SCOPUS:0023660273
SN - 0022-2836
VL - 194
SP - 747
EP - 750
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 4
ER -