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. 1991 Nov 25;19(22):6129–6132. doi: 10.1093/nar/19.22.6129

Mutagenesis of the DNA binding residues in bovine pancreatic DNase 1: an investigation into the mechanism of sequence discrimination by a sequence selective nuclease.

A J Doherty 1, A F Worrall 1, B A Connolly 1
PMCID: PMC329101  PMID: 1956772

Abstract

The sequence selectivity of DNase 1 cleavage has been investigated by site-directed mutagenesis of a chemically synthesised gene. Two key DNA binding residues have been conservatively altered (Y76F and R41K) or have had their side-chains truncated (Y76A and R41A) and the effect on the cleavage of tyr T promoter DNA has been noted. It would appear from these studies that DNase 1 is not sensitive to minor groove width via these DNA-contacting residues, and it is suggested that DNA helical stiffness is a controlling parameter in determining DNase 1 sequence selectivity.

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Selected References

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