Autoregulation of the plasmid addiction operon of bacteriophage P1.
The P1 plasmid addiction operon increases the apparent stability of a plasmid that carries it by killing plasmid-free (cured) segregants. The operon consists of a gene encoding an endotoxin responsible for death on curing (doc), preceded by a gene encoding a relatively unstable antidote that can prevent host death (phd). When the copy number of the operon was increased, expression of a lacZ reporter fused to the promoter of the operon decreased, indicating that expression of the operon was stabilized by an autoregulatory circuit. Transcription of the lacZ reporter was repressed about 10-fold when phd, without doc, was expressed from an exogenous promoter. DNase I footprinting showed that Phd binds a perfect 10-base pair palindromic DNA sequence and, at higher concentrations, an adjacent, imperfect palindrome. The palindromic sites are located between the -10 region of the putative promoter and the start codon of phd. Electrophoretic mobility of DNA containing the promoter region was retarded in the presence of Phd and further retarded in the presence of Phd and Doc. When doc was co-expressed with phd, repression of the lacZ fusion was enhanced more than 100-fold. Thus, both products of the addiction operon participate in its autoregulation.
Magnuson
R
R
Laboratory of Biochemistry, NCI, National Institutes of Health, Bethesda, Maryland 20892-4225, USA.
Lehnherr
H
H
Mukhopadhyay
G
G
Yarmolinsky
M B
MB
D017101:Bacteriophage P1 / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter; D005814:Genes, Viral; D006706:Homeostasis / Q000235:genetics / Q000502:physiology; D007763:Lac Operon; D008969:Molecular Sequence Data; D009876:Operon; D010957:Plasmids / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology
D017931:DNA Primers; D004279:DNA, Viral; C083664:Doc protein, Enterobacteria phage P1; C083665:Phd protein, Enterobacteria phage P1; D014764:Viral Proteins; D003850:Deoxyribonuclease I