The isolated N-terminal DNA binding domain of the c repressor of bacteriophage 16-3 is functional in DNA binding in vivo and in vitro.
The 197 amino acid c repressor of the temperate Rhizobium meliloti phage 16-3 still regulates the OR operator of the phage after removal of its carboxyl terminal region. When cloned in the low-copy-number plasmid pGA46, a severely truncated variant (R1-77), which retains only the first 77 amino acids of the intact protein, repressed in vivo transcription from the phage promoter PR. When the R1-77 repressor was fused to E. coli beta-galactosidase, the hybrid protein bound OR operator DNA in vitro. The behavior of fusion proteins derived from a point mutant is consistent with the assignment of DNA binding specificity to the amino-terminal region. Furthermore two repressor alleles bearing ts mutations that mapped in the R1-77 region (near a helix-turn-helix motif) were also temperature sensitive for regulation of the OR site, while an 18 bp "in frame" deletion mutant, which mapped in the carboxyl terminal segment, regulated the OR operator in wild-type fashion. The carboxyl terminal region of the repressor is however necessary for the control of lysogenic development of 16-3.
Dallmann
G
G
Attila József University, Department of Genetics, Szeged, Hungary.
Marincs
F
F
Papp
P
P
Gaszner
M
M
Orosz
L
L
D000483:Alleles; D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005809:Genes, Regulator; D005814:Genes, Viral; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012231:Rhizobium / Q000235:genetics* / Q000378:metabolism; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D012097:Repressor Proteins