Role of the lytic repressor in prophage induction of phage lambda as analyzed by a module-replacement approach.
Using a module exchange approach, we have tested a long-standing model for the role of Cro repressor in lambda 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(R)3 operator, thereby repressing the cI gene and making the switch irreversible. Here we tested this model by replacing lambda 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 cI, because lac repressor could not repress P(RM) in our constructs. Repression of cI 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.
Department of Biochemistry and Molecular Biophysics, University of Arizona, Life Sciences South Building, 1007 East Lowell Street, Tucson, AZ 85721, USA.
Shota
S
Atsumi
John W
JW
Little
D001426:Bacterial Proteins / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D044127:Epigenesis, Genetic; D015967:Gene Expression Regulation, Viral; D056968:Lac Repressors; D008242:Lysogeny / Q000235:genetics*; D009154:Mutation; D011401:Promoter Regions, Genetic; D039002:Prophages / Q000235:genetics* / Q000502:physiology; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D054334:Viral Regulatory and Accessory Proteins; D014775:Virus Activation / Q000235:genetics*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D056968:Lac Repressors; D012097:Repressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins