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A2 cro, the lysogenic cycle repressor, specifically binds to the genetic switch region of Lactobacillus casei
V Ladero1, P García, J C Alonso
1Facultad de Medicina, Universidad de Oviedo, Julián Clavería s.n., Oviedo, 33006, Spain.
Abstract:
Lysogenic induction of temperate bacteriophage A2 of Lactobacillus casei is controlled by the action of its cI and cro products at the phage operator region. Three 20-bp inverted repeated DNA segments (subsites O1, O2, and O3) and the two divergent (PL and PR) promoters were mapped within the 153-bp operator region. The A2-encoded Cro product is shown to be the functional homolog of lambda Cro. The binding of Cro to the three operator subsites is noncooperative and yields two discrete protein-DNA complexes of retarded migration in mobility shift assays. The Kapp value for the Cro-PL-PR DNA complex was estimated to be 6 nM. Cro shows a slightly higher affinity for O3 than for O1 and O2 subsites. The O3 subsite overlaps the -35 hexamer of the PL promoter, which directs cI expression. A Cro mutant protein, devoid of the last 12 residues (Cro*), allowed the assignment of the DNA-binding domain to the NH2 end of Cro. The C end enhances its affinity for the DNA and probably stabilizes bending induced by Cro.