Kolkhof P, Oehler S, Alex R, Müller-Hill B
Institut für Genetik der Universität zu Köln, Germany.
J Mol Biol. 1995 Mar 31;247(3):396-403. doi: 10.1006/jmbi.1994.0148.
Tetrameric Lac repressor achieves cooperative repression by binding simultaneously to O1 and to one of the auxiliary operators O2 or O3, thereby forcing the intervening DNA into a loop. Dimeric Lac repressor is not able to form DNA loops and consequently shows no cooperative repression. We constructed a dimeric Lac repressor mutant which exhibits increased repression to the lac operon that does not depend on specific operator-repressor-operator loops. This Lac repressor carries a synthetic tail of basic residues attached to its C terminus. With this construct, we observe an increase of the in vivo repression upon addition of auxiliary lac operators to a chromosomal lac operon controlled by O1. This suggests that the basic tail enables dimeric Lac repressor to enhance its repression by additional non-specific DNA contacts.
四聚体乳糖阻遏物通过同时结合O1以及辅助操纵基因O2或O3之一来实现协同阻遏,从而迫使中间的DNA形成环状。二聚体乳糖阻遏物无法形成DNA环,因此不表现出协同阻遏作用。我们构建了一种二聚体乳糖阻遏物突变体,它对乳糖操纵子的阻遏作用增强,且不依赖于特定的操纵基因-阻遏物-操纵基因环。这种乳糖阻遏物在其C末端带有一个由碱性残基组成的合成尾巴。通过这个构建体,我们观察到,在向由O1控制的染色体乳糖操纵子中添加辅助乳糖操纵基因后,体内阻遏作用增强。这表明碱性尾巴使二聚体乳糖阻遏物能够通过额外的非特异性DNA接触来增强其阻遏作用。