Role of the lytic repressor in prophage induction of phage λ as analyzed by a module-replacement approach

摘要

Using a module exchange approach, we have tested a longstanding model for the role of Cro repressor in λ prophage induction. This epigenetic switch from lysogeny to the lytic state occurs on activation of the host SOS system, which leads to specific cleavage of CI repressor. It has been proposed that Cro repressor, which operates during lytic growth and which we shall term the lytic repressor, is crucial to prophage induction. In this view, Cro binds to the O_R3 operator, thereby repressing the cl gene and making the switch irreversible. Here we tested this model by replacing λ Cro with a dimeric form of Lac repressor and adding several lac operators. This approach allowed us to regulate the function of the lytic repressor at will and to prevent it from repressing cl, because lac repressor could not repress P_(RM) in our constructs. Repression of cl by the lytic repressor was not required for prophage induction to occur. However, our evidence suggests that this binding can make induction more efficient, particularly at intermediate levels of DNA damage that otherwise cause induction of only a fraction of the population. These results indicate that this strategy of module exchange will have broad applications for analysis of gene regulatory circuits.