Nucleic Acids Research, 1988, Vol. 16, No. 16 7843-7853
© 1988
Articles |
Mismatch-containing oligonucleotide duplexes bound by the E.coli mutS-encoded protein
Friedrich Miescher Institut PO Box 2543, CH-4002 Basel, Switzerland 1Department of Biochemistry, Duke University Medical Center Durham, NC 27710, USA
Received June 8, 1988. Revised July 1, 1988. Accepted July 26, 1988.
The binding of the mutS gene product, a protein involved in at least two E. coli mismatch correction pathways, to a series of synthetic DNA duplexes containing mismatches or mismatch analogues of the purine/pyrimidine type was studied in order to establish whether a correlation exists between the recognition of these mispairs and the efficiency of their correction in vivo. Experiments using nitrocellulose filter binding or band-shift assays revealed that duplexes containing a G/T mismatch or its analogues I/T and DI/T were bound by the protein with affinities correlating to the efficiency of their repair in vivo. In contrast, the A/C mismatch, contained within the same sequence, was bound only poorly, despite being efficiently corrected in vivo. The analogues of the A/C mispair, uncorrected in vivo, were not detectably bound under the conditions of these assays.
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