J Mol Biol. 1991 Mar 20;218(2):313-21.

The roles of residues 5 and 9 of the recognition helix of Lac repressor in lac operator binding.

Sartorius J, Lehming N, Kisters-Woike B, von Wilcken-Bergmann B, Müller-Hill B.

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Institut für Genetik, Universität zu Köln, Germany.

Abstract

We constructed expression libraries for Lac repressor mutants with amino acid exchanges in positions 1, 2, 5 and 9 of the recognition helix. We then analysed the interactions of residues 5 and 9 with operator variants bearing single or multiple symmetric base-pair exchanges in positions 3, 4 and 5 of the ideal fully symmetric lac operator. We isolated 37 independent Lac repressor mutants with five different amino acids in position 5 of the recognition helix that exhibit a strong preference for particular residues in position 2 and, to a lesser extent, in position 1 of the recognition helix. Our results suggest that residue 5 of the recognition helix (serine 21) contributes to the specific recognition of base-pair 4 of the lac operator. They further suggest that residue 9 of the recognition helix (asparagine 25) interacts non-specifically with a phosphate of the DNA backbone, possibly between base-pairs 2 and 3.

PMID:: 2010911; [PubMed - indexed for MEDLINE]

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